KR102356388B1 - Compositions and methods for modulating angiopoietin-like 3 expression - Google Patents

Abstract

Provided herein are methods, compounds, and compositions for reducing the expression of ANGPTL3 mRNA and protein in an animal. Also provided herein are methods, compounds, and compositions for reducing lipids and/or sugars in an animal. Such methods, compounds, and compositions are useful for treating, preventing, delaying, or ameliorating cardiovascular disease and/or metabolic disease, or any one or more of the symptoms thereof in a subject in need thereof.

Description

COMPOSITIONS AND METHODS FOR MODULATING ANGIOPOIETIN-LIKE 3 EXPRESSION

SEQUENCE LISTING This application has been filed with a Sequence Listing in electronic form. The sequence listing is provided as BIOL0254WOSEQ_ST25.txt (created on Tuesday, April 28, 2015) with a size of 0.98 MB. The information in the electronic form of the Sequence Listing is incorporated herein by reference in its entirety.

field of invention

Provided herein are methods, compounds, and compositions for reducing the expression of angiopoietin-like 3 (ANGPTL3) mRNA and protein in an animal. Also provided herein are methods, compounds, and compositions with an ANGPTL3 inhibitor for reducing an ANGPTL3-related disease or condition in an animal. Such methods, compounds, and compositions are useful for treating, preventing, delaying, or ameliorating any one or more of a cardiovascular or metabolic disease, or symptom thereof, in an animal.

Diabetes and obesity (sometimes collectively referred to as “diabetes due to overweight”) are known to exacerbate the pathology of diabetes and are associated in that more than 60% of diabetic patients are obese. Most human obesity is correlated with insulin resistance and leptin resistance. In fact, it has been suggested that obesity may have a greater effect on insulin action than diabetes itself (Sindelka et al., Physiol Res ., 2002, 51 , 85-91). In addition, several compounds on the market for the treatment of diabetes are known to cause weight gain, which is a highly undesirable side effect for the treatment of this disease.

Cardiovascular disease is also correlated with obesity and diabetes. Cardiovascular disease encompasses multiple etiologies and has an equally wide range of causative agents and interrelated factors. Many causative agents contribute to symptoms such as elevated plasma levels of cholesterol (including non-high-density lipoprotein cholesterol (non-HDL-C)), as well as other lipid-related disorders. Such lipid-related disorders (commonly referred to as dyslipidemia) include, among other indications, hyperlipidemia, hypercholesterolemia and hypertriglyceridemia. Elevated non-HDL cholesterol is associated with atherogenesis and its sequelae, including cardiovascular diseases such as atherosclerosis, coronary artery disease, myocardial infarction, ischemic stroke, and other forms of heart disease. They are named the most common type of disease in industrialized countries. In fact, an estimated 12 million people in the United States have coronary artery disease, and about 36 million require treatment for elevated cholesterol levels.

Epidemiologic and experimental evidence shows that high levels of circulating triglycerides (TG) may be responsible for cardiovascular disease and a myriad of metabolic disorders (Valdivielso et al., 2009, Atherosclerosis Zhang et al., 2008, Circ Res . 1;102(2):250-6). TG derived from exogenous or endogenous sources is incorporated and secreted within chylomicrons from the intestine or in very low density lipoproteins (VLDL) from the liver. Once in circulation, TG is hydrolyzed by lipoprotein lipase (LpL) and the resulting free fatty acids can then be absorbed by local tissues and used as a source of energy. Because of their profound effects, LpL generally has TG and metabolism in plasma, and the discovery and development of compounds that affect LpL activity is of great interest.

Metabolic syndrome is a combination of medical disorders that increase the risk for cardiovascular disease and diabetes. Symptoms, including high blood pressure, high triglycerides, decreased HDL and obesity, coexist in some individuals. It affects large numbers of humans in a cluster-like manner. In some studies, incidence in the United States is calculated as up to 25% of the population. Metabolic syndrome is known by various other names, such as (metabolic) syndrome X, insulin resistance syndrome, Reven's syndrome or CHAOS. With the high incidence of cardiovascular and metabolic disorders, there remains a need for improved approaches to treat these conditions.

Angiopoietins are a family of secreted growth factors. Along with its respective endothelium-specific receptors, angiopoietin plays an important role in angiogenesis. One family member, angiopoietin-like 3 (also known as angiopoietin-like protein 3, ANGPT5, ANGPTL3, or angiopoietin 5), is predominantly expressed in the liver and plays a role in regulating lipid metabolism. (Kaplan et al., J. Lipid Res., 2003 , 44 , 136-143). A whole genome association scan (GWAS) examining the genome for common variants associated with plasma concentrations of HDL, LDL and triglycerides found an association between triglycerides and single-nucleotide polymorphisms (SNPs) proximate to ANGPTL3 (Willer et al.) al., Nature Genetics, 2008, 40(2):161-169). Individuals with allogeneic ANGPTL3 loss-of-function mutations have low levels of atherogenic plasma lipids and lipoproteins, such as total cholesterol (TC) and TG, low-density lipoprotein cholesterol (LDL-C), apoliprotein B (apoB), non -HDL-C, as well as HDL-C (Romeo et al . 2009, J Clin Invest , 119(1):70-79; Musunuru et al . 2010 N Engl J Med , 363:2220-2227; Martin -Campos et al. 2012, Clin Chim Acta , 413:552-555; Minicocci et al. 2012, J Clin Endocrinol Metab , 97:e1266-1275; Noto et al. 2012, Arterioscler Thromb Vasc Biol , 32:805-809; Pisciotta et al. 2012, Circulation Cardiovasc Genet , 5:42-50). This clinical phenotype is termed familial combined hypolipidemia (FHBL2). Despite reduced VLDL secretion, subjects with FHBL2 do not have increased hepatic fat content. They also appear to have lower plasma glucose and insulin levels and, importantly, both diabetes and cardiovascular disease appear to be absent from these subjects. There is no clinically negative phenotype reported to date (Minicocci et al. 2013, J of Lipid Research , 54:3481-3490). Reduction of ANGPTL3 has been shown to result in reductions in TG, cholesterol and LDL levels (US Series No. 13/520,997; PCT Publication WO 2011/085271). ANGPTL3 deficient mice have very low plasma triglyceride (TG) and cholesterol levels, while overexpression has the opposite effect (Koishi et al. 2002; Koster 2005; Fujimoto 2006). Therefore, the potential role of ANGPTL3 in lipid metabolism makes it an attractive target for therapeutic treatment.

To date, therapeutic strategies for treating cardio-metabolic diseases by directly targeting ANGPTL3 levels have been limited. ANGPTL3 polypeptide fragments (US Ser. No. 12/128,545), anti-ANGPTL3 antibodies (US Ser. No. 12/001,012) and ANGPTL3 nucleic acid inhibitors (including antisense oligonucleotides) (US Ser. No. 13/520,997; PCT Publication WO 2011/085271; incorporated herein by reference in its entirety) have previously been proposed or developed, however, certain compounds targeting ANGPTL3 have also not been approved for the treatment of cardiovascular diseases. Therefore, there is an unmet need for highly potent and resistant compounds that inhibit ANGPTL3. The invention disclosed herein relates to the discovery and use of novel, highly potent inhibitors of ANGPTL3 expression in treatment.

Summary of the invention

Provided herein are compositions and methods for modulating expression of ANGPTL3 mRNA and protein. In certain embodiments, the composition is an ANGPTL3 specific inhibitor. In certain embodiments, the ANGPTL3 specific inhibitor reduces the expression of ANGPTL3 mRNA and protein.

In certain embodiments, the composition is an ANGPTL3 specific inhibitor. In certain embodiments, the ANGPTL3 specific inhibitor is a nucleic acid. In certain embodiments, the nucleic acid is an antisense compound. In certain embodiments, the antisense compound is a modified oligonucleotide. In certain embodiments, the antisense compound is a modified oligonucleotide having an attached conjugate group.

In certain embodiments, the ANGPTL3 specific inhibitor is a modified oligonucleotide having a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and at least 8 of the nucleobase sequence of SEQ ID NO: 77 , at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases It has a nucleobase sequence comprising

In certain embodiments, the ANGPTL3 specific inhibitor is a modified oligonucleotide having a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and is equivalent to nucleobases 1140-1159 of SEQ ID NO: 1 and a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases complementary to the length portion, wherein the nucleobase sequence of the modified oligonucleotide is at least 80% complementary to SEQ ID NO:1.

In certain embodiments, the ANGPTL3 specific inhibitor is a modified oligonucleotide having a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and is equivalent to nucleobases 9715-9734 of SEQ ID NO:2 and a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases complementary to the length portion, wherein the nucleobase sequence of the modified oligonucleotide is at least 80% complementary to SEQ ID NO:2.

In certain embodiments, the ANGPTL3 specific inhibitor is a modified oligonucleotide having a conjugate group, wherein the modified oligonucleotide consists of 20 linked nucleosides and comprises at least 8 contiguous nucleobases of SEQ ID NO: 77 and wherein said modified oligonucleotide comprises a nucleobase sequence, said modified oligonucleotide comprising: (a) a gap segment consisting of 10 linked deoxynucleosides; (b) a 5' wing segment consisting of five linked nucleosides; (c) a 3' wing segment consisting of 5 linked nucleosides; wherein said gap segment is located between the 5' wing segment and the 3' wing segment, and each nucleoside of each wing segment comprises a 2'-0-methoxyethyl sugar, wherein each internucleoside linkage This phosphorothioate linkage is where each cytosine residue is 5-methylcytosine.

In certain embodiments, the ANGPTL3 specific inhibitor is a modified oligonucleotide having a conjugate group, wherein the modified oligonucleotide consists of 20 linked nucleosides and a nucleus consisting of at least 8 contiguous nucleobases of SEQ ID NO: 77 nucleotide sequence, wherein the modified oligonucleotide consists of: (a) a gap segment consisting of 10 linked deoxynucleosides; (b) a 5' wing segment consisting of five linked nucleosides; (c) a 3' wing segment consisting of 5 linked nucleosides; wherein said gap segment is located between the 5' wing segment and the 3' wing segment, and each nucleoside of each wing segment comprises a 2'-0-methoxyethyl sugar, wherein each internucleoside linkage This phosphorothioate linkage is where each cytosine residue is 5-methylcytosine.

In certain embodiments, the present disclosure provides conjugated antisense compounds. In certain embodiments, the present disclosure provides a conjugated antisense compound comprising an antisense oligonucleotide complementary to a nucleic acid transcript. In certain embodiments, the present disclosure provides methods of contacting a cell with a conjugated antisense compound comprising an antisense oligonucleotide complementary to a nucleic acid transcript. In certain embodiments, the present disclosure provides methods of contacting a cell with a conjugated antisense compound comprising an antisense oligonucleotide and reducing the amount or activity of a nucleic acid transcript in a cell.

The asialoglycoprotein receptor (ASGP-R) has been previously described. See, eg, Park et al., PNAS vol. 102, No. 47, pp 17125-17129 (2005). Such receptors are expressed on liver cells, particularly hepatocytes. Furthermore, it has been shown that compounds comprising a cluster of three N-acetylgalactosamine (GalNAc) ligands can bind ASGP-Rs, resulting in uptake of these compounds into cells. See, eg, Khorev et al., Bioorganic and Medicinal Chemistry, 16, 9, pp 5216-5231 (May 2008). Accordingly, conjugates containing such a GalNAc cluster have been used to promote the uptake of certain compounds into hepatocytes, specifically hepatocytes. For example, certain GalNAc-containing conjugates have been shown to increase the activity of double-stranded siRNA compounds in liver cells in vivo. In such cases, the GalNAc-containing conjugate is typically attached to the sense strand of the siRNA duplex. Since the sense strand is discarded before the antisense strand eventually hybridizes with the target nucleic acid, the conjugate is unlikely to interfere with activity. Typically, the conjugate is attached to the 3' end of the sense strand of the siRNA. For example, U.S. See Patent No. 8,106,022. Certain conjugate groups described herein are more active and/or easier to synthesize than those previously described conjugate groups.

In certain embodiments of the invention, conjugates are attached to single-stranded antisense compounds, including, but not limited to, RNase H based antisense compounds and antisense compounds that alter splicing of a target nucleic acid of a pre-mRNA. In such embodiments, the conjugate must remain attached to the antisense compound long enough to provide the benefit (improved uptake into the cell), but is subsequently cleaved, or hybridized and spliced to the target nucleic acid, or It may not interfere with subsequent steps required for activity, such as interactions with RNase H or enzymes involved in splice regulation. This balance of properties is more important in the setting of single-stranded antisense compounds than in siRNA compounds, and the conjugate can be conveniently attached to the sense strand. Disclosed herein are conjugated single-stranded antisense compounds that have improved efficacy in liver cells in vivo compared to the same antisense compound lacking the conjugate. Such improved efficacy is surprising given the required balance of properties for these compounds.

In certain embodiments, a conjugate group herein comprises a cleavable moiety. As mentioned, without being bound by mechanism, it is logical that the conjugate should remain on the compound long enough to provide absorption enhancement, but then, some or ideally all of the conjugate is cleaved to It is desirable to release the parent compound (eg, antisense compound) in its most active form. In certain embodiments, the cleavable moiety is a cleavable nucleoside. Such embodiments take advantage of endogenous nucleases in the cell by attaching the remainder of the conjugate (cluster) to the antisense oligonucleotide via a nucleoside via one or more cleavable bonds, such as phosphodiester linkages. In certain embodiments, the cluster is linked to the cleavable nucleoside via a phosphodiester linkage. In certain embodiments, the cleavable nucleoside is attached to the antisense oligonucleotide (antisense compound) by a phosphodiester linkage. In certain embodiments, a conjugate group may contain 2 or 3 cleavable nucleosides. In such embodiments, such cleavable nucleosides are linked to each other and/or linked to an antisense compound and/or linked to a cluster via a cleavable bond (eg, a phosphodiester linkage). Certain conjugates herein do not contain a cleavable nucleoside but instead contain a cleavable bond. It appears that sufficient cleavage of the conjugate from the oligonucleotide is provided by at least one bond that is susceptible to cleavage in the cell (cleavable bond).

In certain embodiments, the conjugated antisense compound is a prodrug. Such prodrugs are administered to the animal and eventually metabolized to a more active form. For example, a conjugated antisense compound is cleaved to remove all or part of the conjugate, resulting in an active (or more active) form of the antisense compound lacking all or part of the conjugate.

In certain embodiments, the conjugate is attached at the 5' end of the oligonucleotide. Certain such 5'-conjugates are cleaved more efficiently than their counterparts with similar conjugate groups attached at the 3' end. In certain embodiments, improved activity may be associated with improved cleavage. In certain embodiments, oligonucleotides comprising a conjugate at the 5' end have greater potency than oligonucleotides comprising a conjugate at the 3' end (see, e.g., Examples 56, 81, 83, and 84). ). Moreover, the 5'-attachment allows for simpler oligonucleotide synthesis. Typically, oligonucleotides are synthesized on a solid support in the 3' to 5' direction. To prepare 3'-conjugated oligonucleotides, typically pre-conjugated 3' nucleosides are attached to a solid support followed by conventional preparation of the oligonucleotides. However, attachment of the conjugated nucleoside to a solid support is problematic for synthesis. Moreover, using this approach can limit the types of reactions and reagents that can then be present throughout the synthesis of the oligonucleotide and degraded or used during subsequent steps of the conjugate. Using the structures and techniques described herein for 5'-conjugated oligonucleotides, one skilled in the art can synthesize the oligonucleotide using standard automated techniques and the final (5'-most end) after the oligonucleotide is cleaved from the solid support. ) conjugates with nucleosides can be introduced.

In light of the art and this disclosure, one of ordinary skill in the art can readily prepare any of the conjugates and conjugated oligonucleotides herein. Moreover, the synthesis of certain such conjugates and conjugated oligonucleotides disclosed herein is easier and/or requires few steps and is therefore less expensive than the previously disclosed conjugates, providing advantages in manufacturing. For example, the synthesis of specific conjugate groups consists of fewer synthetic steps, resulting in increased yields compared to the previously described conjugate groups. Conjugate groups such as GalNAc3-10 in Example 46 and GalNAc3-7 in Example 48 require the assembly of more chemical intermediates. 8,106,022 or U.S. It is much simpler than the previously described conjugates such as those described in 7,262,177. Accordingly, these and other conjugates described herein have advantages over previously described compounds for use with any oligonucleotide, including either single-stranded oligonucleotides or double-stranded oligonucleotides (e.g., siRNA). have

Similarly, groups of conjugates having only one or two GalNAc ligands are disclosed herein. As can be seen, such conjugate groups improve the activity of antisense compounds. Such compounds are much easier to prepare than conjugates comprising three GalNAc ligands. Conjugate groups comprising one or two GalNAc ligands can be attached to any antisense compound, including either single-stranded oligonucleotides and double-stranded oligonucleotides (eg, siRNA).

In certain embodiments, the conjugates herein do not substantially change a particular measure of resistance. For example, conjugated antisense compounds are shown herein to be no more immunogenic than the unconjugated parent compound. As the efficacy improves, embodiments in which the tolerance remains the same (or in fact the tolerance is only slightly deteriorated compared to the increased efficacy) has improved properties for therapy.

In certain embodiments, conjugation allows to alter the antisense compound in a way that has less attractive results in the absence of conjugation. For example, in certain embodiments, completely replacing one or more phosphorothioate linkages of a phosphorothioate antisense compound with a phosphodiester linkage results in an improvement in some measure of resistance. For example, in certain instances, such antisense compounds having one or more phosphodiesters are less immunogenic than the same compound in which each linkage is a phosphorothioate. However, in certain instances, the same replacement of one or more phosphorothioate linkages with phosphodiester linkages, as shown in Example 26, also results in reduced cellular uptake and/or loss of potency. In certain embodiments, the conjugated antisense compounds described herein withstand such changes in linkage with little or no loss in absorption and potency compared to their conjugated full-phosphorothioate counterparts. Indeed, in certain embodiments, for example, in Examples 44, 57, 59, and 86, the conjugates and oligonucleotides comprising at least one phosphodiester internucleoside linkage actually also comprise the same conjugate. showed increased potency in vivo even compared to its full phosphorothioate counterpart. Moreover, as conjugation results in a substantial increase in absorption/potency, a small loss in this substantial increase may be acceptable to achieve improved tolerability. Thus, in certain embodiments, the conjugated antisense compound comprises at least one phosphodiester linkage.

In certain embodiments, conjugation of an antisense compound herein results in increased delivery, uptake and activity in hepatocytes. Thus, more compounds are delivered to the liver tissue. However, in certain embodiments, such increased delivery alone does not account for the overall increase in activity. In certain such embodiments, more of the compound enters the hepatocytes. In certain embodiments, even increased hepatocyte uptake does not account for the overall increase in activity. In such embodiments, the productive absorption of the conjugated compound is increased. For example, as shown in Example 102, certain embodiments of GalNAc-containing conjugates increase the enrichment of antisense oligonucleotides in hepatocytes compared to non-parenchymal cells. This enrichment is beneficial for oligonucleotides targeting genes expressed in hepatocytes.

In certain embodiments, the conjugated antisense compounds herein result in reduced renal exposure. For example, as shown in Example 20, the concentration of antisense oligonucleotides comprising certain embodiments of GalNAc-containing conjugates is lower in the kidney than antisense oligonucleotides without GalNAc-containing conjugates. It has several beneficial therapeutic implications. For therapeutic indications where activity in the kidney is not sought, exposure to the kidney jeopardizes renal toxicity without a corresponding benefit. In addition, high concentrations in the kidneys typically cause a loss of the compound into the urine, thereby gaining faster clearance. Thus, for non-renal targets, renal accumulation is undesirable.

In certain embodiments, the present disclosure provides a conjugated antisense compound represented by the formula:

here,

A is an antisense oligonucleotide;

B is a cleavable moiety

C is a conjugate linker

D is a branching group

each E is a linking group;

each F is a ligand; and

q is an integer from 1 to 5;

In the diagrams above and similar diagrams herein, the branching group “D” is branched as many times as necessary to accommodate the number of (E-F) groups as indicated by “q”. Thus, when q = 1, the formula is:

When q = 2, the formula is:

When q = 3, the formula is:

When q = 4, the formula is:

When q = 5, the formula is:

In certain embodiments, the conjugated antisense compound is provided having the structure:

.

.

In certain embodiments, the conjugated antisense compound is provided having the structure:

.

.

In certain embodiments, the conjugated antisense compound is provided having the structure:

.

.

In certain embodiments, the conjugated antisense compound is provided having the structure:

In embodiments having more than one (eg, more than one "m" or "n") of a particular variable, unless otherwise indicated, each such particular variable is independently selected. Thus, for structures with n greater than 1, each n is selected independently, which may or may not be the same as each other.

In certain embodiments, the present disclosure provides a conjugated antisense compound represented by the structure: In certain embodiments, the antisense compound comprises a modified oligonucleotide ISIS 563580 having a 5'-X, wherein X is a conjugate group comprising GalNAc. In certain embodiments, the antisense compound consists of a modified oligonucleotide ISIS 563580 having a 5'-X, wherein X is a conjugate group comprising GalNAc.

In certain embodiments, the present disclosure provides a conjugated antisense compound represented by the structure: In certain embodiments, the antisense compound comprises a conjugated modified oligonucleotide ISIS 703801. In certain embodiments, the antisense compound consists of a conjugated modified oligonucleotide ISIS 703801.

In certain embodiments, the present disclosure provides a conjugated antisense compound represented by the structure: In certain embodiments, the antisense compound comprises a conjugated modified oligonucleotide ISIS 703802. In certain embodiments, the antisense compound consists of a conjugated modified oligonucleotide ISIS 703802.

In certain embodiments, the present disclosure provides a conjugated antisense compound represented by the structure: In certain embodiments, the antisense compound comprises a modified oligonucleotide having the nucleobase sequence of SEQ ID NO: 77, which has 5'-GalNAc with variability in the sugar modifications of the wing. In certain embodiments, the antisense compound consists of a modified oligonucleotide having the nucleobase sequence of SEQ ID NO: 77, which has 5'-GalNAc with variability in the sugar modifications of the wing.

wherein R ¹ is —OCH ₂ CH ₂ OCH ₃ (MOE) and R ² is H; or R ¹ and R ² together form a bridge, wherein R ¹ is —O— and R ² is —CH ₂ —, —CH(CH ₃ )—, or —CH ₂ CH ₂ —, and R ¹ and R ² is directly linked, and the bridge thereby obtained is selected from: -O-CH ₂ -, -O-CH(CH ₃ )-, and -O-CH ₂ CH ₂ -;

^{and for each pair of R 3} and R ⁴ on the same ring, independently for each ring: either R ³ is selected from H and —OCH ₂ CH ₂ OCH ₃ ^{and R 4} is H; or R ³ and R ⁴ together form a bridge, wherein R ³ is —O—, and R ⁴ is CH ₂ —, —CH(CH ₃ )—, or —CH ₂ CH ₂ —, R ³ and R ⁴ is directly linked, and the bridge thereby obtained is selected from: -O-CH ₂ -, -O-CH(CH ₃ )-, and -O-CH ₂ CH ₂ -;

and R ⁵ is selected from H and —CH _{3 ;}

and Z is selected from ^{S -} and O ^-.

Certain embodiments provide a composition comprising a conjugated antisense compound described herein, or a salt thereof, and a pharmaceutically acceptable carrier or diluent.

In certain embodiments, modulation of ANGPTL3 expression occurs in a cell or tissue. In certain embodiments, the modulation occurs in a cell or tissue of an animal. In certain embodiments, the animal is a human. In certain embodiments, the modulation is a decrease in ANGPTL3 mRNA levels. In certain embodiments, the modulation is a decrease in the ANGPTL3 protein level. In certain embodiments, both ANGPTL3 mRNA and protein levels are reduced. Such reduction may occur in a time-dependent manner or in a dose-dependent manner.

Certain embodiments provide compositions and methods for use in therapy. Certain embodiments provide compositions and methods for preventing, treating, delaying, delaying the progression, and/or ameliorating ANGPTL3-related diseases, disorders, and conditions. In certain embodiments, such diseases, disorders, and conditions are cardiovascular and/or metabolic diseases, disorders, and conditions. In certain embodiments, compositions and methods for therapy comprise administering to a subject in need thereof an ANGPTL3 specific inhibitor. In certain embodiments, the ANGPTL3 specific inhibitor is a nucleic acid. In certain embodiments, the nucleic acid is an antisense compound. In certain embodiments, the antisense compound is a modified oligonucleotide. In certain embodiments, the antisense compound is a modified oligonucleotide having an attached conjugate group.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only, and do not limit the invention as claimed. As used herein, the use of the singular includes the plural unless specifically stated otherwise. As used herein, the use of “or” means “and/or” unless stated otherwise. Moreover, use of the term "comprising" as well as other forms such as "comprises" and "included" is not limiting. Also, terms such as "element" or "component" include both elements and components comprising one unit and elements and components comprising more than one subunit, unless specifically stated otherwise.

Section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described. All documents or portions of documents cited in this application, including but not limited to patents, patent applications, articles, books, and articles, are expressly incorporated by reference in their entirety for any purpose.

Justice

Unless specific definitions are provided, the nomenclature employed in connection with, and procedures and techniques for, analytical chemistry, synthetic organic chemistry, and medical and pharmaceutical chemistry described herein refer to those well known and commonly used in the art. to be. Standard techniques can be used for chemical synthesis and chemical analysis. Certain such techniques and procedures can be found, for example, in "Carbohydrate Modifications in Antisense Research" Edited by Sangvi and Cook, American Chemical Society, Washington DC, 1994; "Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, Pa., 21 ^st edition, 2005; and “Antisense Drug Technology, Principles, Strategies, and Applications” Edited by Stanley T. Crooke, CRC Press, Boca Raton, Florida; And Sambrook et al., "Molecular Cloning , A laboratory Manual," 2 nd Edition, Cold Spring Harbor Laboratory Press, 1989 ( which may be incorporated herein by reference for any purpose). Where permitted, all documents, or portions of documents, cited in this application, including, but not limited to, all patents, patent applications, published patent applications and other journal publications, GENBANK Accession numbers and related sequence information obtainable through databases such as the National Center for Biotechnology Information (NCBI) and other data referred to in its entirety herein, as well as in its entirety, are part of the documents discussed herein. incorporated by reference to

Unless otherwise indicated, the following terms have the following meanings:

As used herein, “nucleoside” refers to a compound comprising a nucleobase moiety and a sugar moiety. Nucleosides include, but are not limited to: naturally occurring nucleosides (found in DNA and RNA) and modified nucleosides. The nucleoside may be linked to a phosphate moiety.

As used herein, "chemical modification" refers to a chemical difference in a compound as compared to its naturally occurring counterpart. Chemical modifications of oligonucleotides include nucleoside modifications (including sugar moiety modifications and nucleobase modifications) and internucleoside linkage modifications. With respect to oligonucleotides, chemical modifications do not involve only differences in nucleobase sequence.

As used herein, "furanosyl" means a structure comprising a 5-membered ring comprising four carbon atoms and one oxygen atom.

As used herein, "naturally occurring sugar moiety" means either ribofuranosyl found in naturally occurring RNA or deoxyribofuranosyl found as naturally occurring DNA.

As used herein, "sugar moiety" refers to a naturally occurring sugar moiety or a modified sugar moiety of a nucleoside.

As used herein, "modified sugar moiety" refers to a substituted sugar moiety or sugar surrogate.

As used herein, "substituted sugar moiety" means a furanosyl that is not a naturally occurring sugar moiety. Substituted sugar moieties include, but are not limited to, furanosyl containing substituents at the 2'-position, 3'-position, 5'-position and/or 4'-position. Certain substituted sugar moieties are bicyclic sugar moieties.

As used herein, "2'-substituted sugar moiety" means a furanosyl comprising a substituent at the 2'-position other than H or OH. Unless otherwise indicated, a 2'-substituted sugar moiety is not a bicyclic sugar moiety (i.e., a 2'-substituent of a 2'-substituted sugar moiety does not form a bridge to another atom of the furanosyl ring. does not

As used herein, “MOE” means —OCH ₂ CH ₂ OCH ₃ .

As used herein, "2'-F nucleoside" means a nucleoside comprising a sugar comprising a fluorine in the 2' position. Unless otherwise indicated, the fluorine in the 2'-F nucleoside is at the ribo position (replacing the OH of the native ribose).

The term "sugar surrogate" as used herein refers to a structure that does not contain a furanosyl and is capable of replacing the naturally occurring sugar moiety of a nucleoside, whereby the resulting nucleoside sub-units are to be linked together. and/or linked to other nucleosides to form oligomeric compounds capable of hybridizing complementary oligomeric compounds. Such structures include rings containing atoms different than furanosyl (eg, 4, 6, or 7-membered rings); replacement of the oxygen of the furanosyl by a non-oxygen atom (eg, carbon, sulfur, or nitrogen); or both a change in the number of atoms and replacement of oxygen. Such structures may also include substitutions corresponding to those in place of a substituted sugar moiety (eg, a 6-membered carbocyclic bicyclic sugar surrogate optionally comprising additional substituents). Sugar substitutes also include more complex sugar substitutes (eg, non-ring systems of peptide nucleic acids). Sugar substitutes include, but are not limited to, morpholino, cyclohexenyl and cyclohexitol.

As used herein, a “bicyclic sugar moiety” includes a bridge (including but not limited to furanosyl) that connects two atoms of a 4 to 7 membered ring to form a second ring, resulting in a bicyclic structure. ) means a modified sugar moiety comprising a 4 to 7 membered ring. In certain embodiments, the 4-7 membered ring is a sugar ring. In certain embodiments the 4 to 7 membered ring is furanosyl. In certain such embodiments, a bridge connects the 2'-carbon and the 4'-carbon of the furanosyl.

As used herein, “nucleotide” refers to a nucleoside further comprising a phosphate linking group. As used herein, “linked nucleosides” may or may not be linked by phosphate linkages and thus include, but are not limited to, “linked nucleotides”. As used herein, a “linked nucleoside” is a nucleoside linked in a contiguous sequence (ie, no additional nucleosides are present between the linked ones).

As used herein, "nucleobase" means a group of atoms that can be linked to a sugar moiety to make a nucleoside that can be incorporated into an oligonucleotide, said group of atoms being another oligonucleotide or nucleic acid It can bind to complementary naturally occurring nucleobases. Nucleobases may be naturally occurring or may be modified. "Nucleobase sequence" means the sequence of contiguous nucleobases independent of any sugars, linkages and/or nucleobase modifications.

As used herein, the terms "unmodified nucleobase" or "naturally occurring nucleobase" refer to the naturally occurring heterocyclic nucleobases of RNA or DNA: the purines adenine (A) and guanine (G), and pyrimi thymine (T), cytosine (C) (including 5-methyl C), and uracil (U).

As used herein, "modified nucleobase" means any nucleobase that is not a naturally occurring nucleobase.

As used herein, "modified nucleoside" refers to a nucleoside comprising at least one chemical modification compared to a naturally occurring RNA or DNA nucleoside. Modified nucleosides include modified sugar moieties and/or modified nucleobases.

As used herein, “bicyclic nucleoside” or “BNA” refers to a nucleoside comprising a bicyclic sugar moiety.

As used herein, "restricted ethyl nucleoside" or "cEt" means a nucleoside comprising a bicyclic sugar moiety comprising a _{4'-CH(CH 3 )-O-2' bridge.}

As used herein, "locked nucleic acid nucleoside" or "LNA" refers to a nucleoside comprising a bicyclic sugar moiety comprising a _{4'-CH 2 -O-2' bridge.}

As used herein, "2'-substituted nucleoside" means a nucleoside comprising a substituent at the 2'-position other than H or OH. Unless otherwise indicated, a 2'-substituted nucleoside is not a bicyclic nucleoside.

As used herein, "deoxynucleoside" means a nucleoside comprising a 2'-H furanosyl sugar moiety, as found in naturally occurring deoxyribonucleosides (DNA). . In certain embodiments, the 2'-deoxynucleoside may comprise a modified nucleobase or may comprise an RNA nucleobase (eg, uracil).

As used herein, "oligonucleotide" refers to a compound comprising a plurality of linked nucleosides. In certain embodiments, the oligonucleotide comprises one or more unmodified ribonucleosides (RNA) and/or unmodified deoxyribonucleosides (DNA) and/or one or more modified nucleosides.

"Oligonucleoside" as used herein refers to an oligonucleotide wherein none of the internucleoside linkages contain a phosphorus atom. As used herein, oligonucleotides include oligonucleosides.

As used herein, "modified oligonucleotide" means an oligonucleotide comprising at least one modified nucleoside and/or at least one modified internucleoside linkage.

As used herein, “linkage” or “linking group” refers to a group of atoms that are linked together with two or more other groups of atoms.

"Internucleoside linkage" as used herein refers to a covalent bond between adjacent nucleosides in an oligonucleotide.

"Naturally occurring internucleoside linkage" as used herein refers to a 3' to 5' phosphodiester linkage.

As used herein, "modified internucleoside linkage" means any internucleoside linkage other than a naturally occurring internucleoside linkage.

As used herein, “terminal internucleoside linkage” refers to a linkage between the last two nucleosides of an oligonucleotide or defined region thereof.

As used herein, "phosphorus linking group" means a linking group comprising a phosphorus atom. Phosphorus linking groups include, but are not limited to, groups having the formula:

In the above formula,

each of R _a and R _d is, independently, O, S, CH ₂ , NH, or NJ ₁ , wherein J ₁ is C ₁ -C ₆ alkyl or substituted C ₁ -C ₆ alkyl;

R _b is O or S;

R _c is OH, SH, C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted C ₁ -C ₆ alkoxy, amino or substituted amino; and

J ₁ is R _b is O or S.

Phosphorus linking groups include, but are not limited to, phosphodiesters, phosphorothioates, phosphorodithioates, phosphonates, phosphoramidates, phosphorothioamidates, thioalkylphosphonates, phosphotri esters, thionoalkylphosphotriesters and boranophosphates.

As used herein, "internucleoside phosphorus linking group" means a phosphorus linking group that directly connects two nucleosides.

As used herein, “non-internucleoside phosphorus linking group” refers to a phosphorus linking group that does not directly link two nucleosides. In certain embodiments, a non-internucleoside phosphorus linking group links the nucleoside to a group other than the nucleoside. In certain embodiments, a non-internucleoside phosphorus linking group connects two groups, certain of which are not nucleosides.

As used herein, "neutral linking group" means an uncharged linking group. Neutral linking groups include, but are not limited to: phosphotriesters, methylphosphonates, MMI (-CH ₂ -N(CH ₃ )-O-), amide-3 (-CH ₂ -C(=O)- N(H)-), amide-4 (-CH ₂ -N(H)-C(=O)-), formacetal (-O-CH ₂ -O-), and thioformacetal (-S-CH ₂ -O-). Additional neutral linking groups include nonionic linkages including siloxanes (dialkylsiloxanes), carboxylate esters, carboxamides, sulfides, sulfonate esters and amides (see for example: Carbohydrate Modifications in Antisense Research; YS Sanghvi and PD Cook Eds. ACS Symposium Series 580; Chapters 3 and 4, (pp. 40-65)). Additional neutral linking groups include nonionic linkages comprising mixed N, O, S and CH _{2 component moieties.}

As used herein, "internucleoside neutral linking group" means a neutral linking group that directly connects two nucleosides.

As used herein, "non-internucleoside neutral linking group" refers to a neutral linking group that does not directly link two nucleosides. In certain embodiments, a non-internucleoside neutral linking group links the nucleoside to a group other than the nucleoside. In certain embodiments, a non-internucleoside neutral linking group connects two groups, neither of which is a nucleoside.

As used herein, "oligomeric compound" refers to a polymeric structure comprising two or more substructures. In certain embodiments, the oligomeric compound comprises an oligonucleotide. In certain embodiments, the oligomeric compound comprises one or more conjugate groups and/or end groups. In certain embodiments, the oligomeric compound consists of oligonucleotides. Oligomeric compounds also include naturally occurring nucleic acids. In certain embodiments, the oligomeric compound comprises a backbone of one or more linked monomeric subunits, wherein each linked monomeric subunit is attached directly or indirectly to a heterocyclic base moiety. In certain embodiments, the oligomeric compound may also include monomeric subunits that are not linked to a heterocyclic base moiety, thereby providing an abasic moiety. In certain embodiments, the linkage connecting the monomer subunit, sugar moiety or surrogate and the heterocyclic base moiety can be independently modified. In certain embodiments, linkage-sugar units, which may or may not include heterocyclic bases, may be substituted with a monomer in a mimic such as a peptide nucleic acid.

As used herein, "end group" means one or more atoms attached to one or both of the 3' end or the 5' end of an oligonucleotide. In certain embodiments the terminal group is a conjugate group. In certain embodiments, the terminal group comprises one or more terminal group nucleosides.

As used herein, "conjugate" or "conjugate group" refers to an atom or group of atoms attached to an oligonucleotide or oligomeric compound. In general, the conjugate group modifies one or more properties of the attached compound, including, but not limited to, pharmacokinetic, pharmacodynamic, binding, uptake, cellular distribution, cellular uptake, charge and/or clearance properties.

As used herein, "conjugate linker" or "linker" in the context of a group of conjugates includes any atom or group of atoms and includes (1) an oligonucleotide to another portion of a group of conjugates or (2) ) means the distinction of a conjugate group that is covalently bonded to two or more parts of the conjugate group.

A conjugate group is shown herein as a radical that provides a bond that forms a covalent bond to an oligomeric compound such as an antisense oligonucleotide. In certain embodiments, the point of attachment on the oligomeric compound is the 3'-oxygen atom of the 3'-hydroxyl group of the 3' terminal nucleoside of the oligomeric compound. In certain embodiments the point of attachment on the oligomeric compound is the 5'-oxygen atom of the 5'-hydroxyl group of the 5' terminal nucleoside of the oligomeric compound. In certain embodiments, the bond forming the attachment to the oligomeric compound is a cleavable bond. In certain such embodiments, such cleavable linkages constitute all or part of a cleavable moiety.

In certain embodiments, a conjugate group comprises a cleavable moiety (eg, a cleavable bond or a cleavable nucleoside) and a carbohydrate cluster moiety, such as a GalNAc cluster moiety. Such carbohydrate cluster moieties include: a targeting moiety, and optionally, a conjugate linker. In certain embodiments, carbohydrate cluster moieties are identified by the number and identity of ligands. For example, in certain embodiments, the carbohydrate cluster portion comprises three GalNAc groups and is termed _{“GalNAc 3 ”.} In certain embodiments, the carbohydrate cluster portion comprises four GalNAc groups and is termed _{“GalNAc 4 ”.} Specific carbohydrate cluster moieties (with specific linkers, branching and conjugate linker groups) are described herein and are designated by a roman numeral followed by the subscript “a”. Thus "GalNac3-1 _a " refers to a conjugate group having three GalNAc groups and a specific carbohydrate cluster portion of the specifically identified linking group, branching and linking group. Such carbohydrate cluster fragments are attached to the oligomeric compound via a cleavable moiety, such as a cleavable bond or a cleavable nucleoside.

As used herein, "cleavable moiety" refers to a bond or group capable of being cleaved under physiological conditions. In certain embodiments, the cleavable moiety is cleaved within a cell or sub-cellular compartment, such as a lysosome. In certain embodiments, the cleavable moiety is cleaved by an endogenous enzyme, such as a nuclease. In certain embodiments, a cleavable moiety comprises a group of atoms having 1, 2, 3, 4, or more 4 cleavable bonds.

As used herein, "cleavable bond" means any chemical bond that can be cleaved. In certain embodiments, the cleavable bond is selected from: an amide, a polyamide, an ester, an ether, an ester of a phosphodiester, or both, a phosphate ester, a carbamate, a di-sulfide, or a peptide.

As used herein, "carbohydrate cluster" refers to a compound having one or more carbohydrate moieties attached to a scaffold or linker group. For example the following (Reference: Maier et al, "Synthesis of Antisense Oligonucleotides Conjugated to a Multivalent Carbohydrate Cluster for Cellular Targeting," Bioconjugate Chemistry, 2003, (14):. 18-29, this is incorporated herein in its entirety by reference , or Rensen et al., "Design and Synthesis of Novel N- Acetylgalactosamine-Terminated Glycolipids for Targeting of Lipoproteins to the Hepatic Asiaglycoprotein Receptor," J. Med . Chem . 2004, (47): 5798-5808 (Carbohydrate Conjugates) As an example of a gate cluster)).

As used herein, "modified carbohydrate" means any carbohydrate that has one or more chemical modifications to a naturally occurring carbohydrate.

As used herein, "carbohydrate derivative" means any compound that can be synthesized using a carbohydrate as a starting material or intermediate.

As used herein, “carbohydrate” refers to a naturally occurring carbohydrate, a modified carbohydrate, or a carbohydrate derivative.

"Protecting group" means any compound or protecting group known to one of ordinary skill in the art. Non-limiting examples of protecting groups can be found in "Protective Groups in Organic Chemistry", TW Greene, PGM Wuts , ISBN 0-471-62301-6, John Wiley & Sons, Inc, New York (in its entirety). incorporated herein by reference).

As used herein, “single-stranded” refers to an oligomeric compound that does not hybridize to its complement and lacks sufficient self-complementarity to form a stable self-duplex. As used herein, “double-stranded” refers to a pair of oligomeric compounds that are hybridized to each other or a single self-complementary oligomeric compound that forms a hairpin structure.

In certain embodiments, the double-stranded oligomeric compound comprises a first and a second oligomeric compound. As used herein, "antisense compound" means a compound comprising or consisting of at least a portion of an oligonucleotide complementary to a hybridizable target nucleic acid, wherein at least one antisense activity occurs.

As used herein, "antisense activity" means any detectable and/or measurable change attributable to hybridization of an antisense compound to its target nucleic acid. In certain embodiments, antisense activity comprises modulating the amount or activity of a target nucleic acid transcript (eg, mRNA). In certain embodiments, antisense activity comprises modulation of splicing of pre-mRNA.

As used herein, “RNase H-based antisense compound” refers to an antisense compound, wherein at least a portion of the antisense activity of the antisense compound is derived from hybridization of the antisense compound to the target nucleic acid and subsequent RNase H of the target nucleic acid. Amputation is the cause.

As used herein, “RISC-based antisense compound” refers to an antisense compound, wherein at least a portion of the antisense activity of the antisense compound is attributable to the RNA-induced silencing complex (RISC).

As used herein, “detection” or “measurement” means that a test or assay for detection or measurement is performed. Such detection and/or measurement may result in a value of zero. Thus, if a test for detection or measurement expressed no activity (zero activity), a step of detecting or measuring said activity was nevertheless performed.

As used herein, "detectable and/or measurable activity" means a non-zero statistically significant activity.

As used herein, "essentially unchanged" means little or no change in a particular parameter, particularly with respect to another parameter that varies even more. In certain embodiments, a parameter is essentially unchanged when it changes less than 5%. In certain embodiments, a parameter changes less than 2-fold but is essentially unchanged if another parameter changes at least 10-fold. For example, in certain embodiments, antisense activity is a change in the amount of a target nucleic acid. In certain such embodiments, the amount of non-target nucleic acid is essentially unchanged, provided that it varies much less than the target nucleic acid, but the change need not be zero.

As used herein, “expression” refers to the process by which a gene eventually becomes a protein. Expression includes, but is not limited to, transcription, post-transcriptional modifications (eg, splicing of the 5'-cap, polyadenylation, addition), and translation.

As used herein, “target nucleic acid” refers to a nucleic acid molecule intended to hybridize to an antisense compound to produce the desired antisense activity. An antisense oligonucleotide has sufficient complementarity to its target nucleic acid to allow hybridization under physiological conditions.

As used herein, "nucleobase complementarity" or "complementarity," when referring to a nucleobase, refers to a nucleobase capable of base pairing with another nucleobase. For example, in DNA, adenine (A) is complementary to thymine (T). For example, in RNA, adenine (A) is complementary to uracil (U). In certain embodiments, a complementary nucleobase refers to a nucleobase of an antisense compound capable of base pairing with a nucleobase of its target nucleic acid. For example, if a nucleobase at a specific position in an antisense compound is capable of hydrogen bonding with a nucleobase at a specific position in the target nucleic acid, then the position of hydrogen bonding between the oligonucleotide and the target nucleic acid is considered complementary in the nucleobase pair. . A nucleobase comprising a particular modification may retain the ability to pair with its counterpart nucleobase and thus still maintain nucleobase complementarity.

As used herein, "non-complementary" with respect to a nucleobase means a pair of nucleobases that do not form hydrogen bonds with each other.

As used herein, "complementary" in reference to an oligomeric compound (eg, linked nucleoside, oligonucleotide, or nucleic acid) means hybridizing to another oligomeric compound or a region thereof through nucleobase complementarity. for such an oligomeric compound or a region thereof. Complementary oligomeric compounds need not have nucleobase complementarity at each nucleoside. Rather, some mismatch is tolerated. In certain embodiments, the complementary oligomeric compound or region is complementary at 70% of the nucleobases (70% complementary). In certain embodiments, complementary oligomeric compounds or regions are 80% complementary. In certain embodiments, complementary oligomeric compounds or regions are 90% complementary. In certain embodiments, complementary oligomeric compounds or regions are 95% complementary. In certain embodiments, complementary oligomeric compounds or regions are 100% complementary.

As used herein, "mismatch" means a nucleobase of a first oligomeric compound that is unable to pair with a nucleobase at the corresponding position of the second oligomeric compound when the first and second oligomeric compounds are aligned. . One or both of the first and second oligomeric compounds may be oligonucleotides.

As used herein, "hybridization" refers to the pairing of a complementary oligomeric compound (eg, an antisense compound and its target nucleic acid). Without being limited to a particular mechanism, the most common mechanism of mating involves hydrogen bonding, which can be Watson-Crick, Hugsteen, or inverted Hugsteen hydrogen bonding between complementary nucleobases.

As used herein, "specifically hybridizes" refers to the ability of an oligomeric compound to hybridize to one nucleic acid site with greater affinity than it hybridizes to another nucleic acid site.

As used herein, "fully complementary" to an oligonucleotide or portion thereof means that each nucleobase of the oligonucleotide or portion thereof is capable of pairing with a complementary nucleic acid or a nucleobase of its contiguous region. it means. Thus, a fully complementary region does not contain mismatched or unhybridized nucleobases in one strand.

As used herein, "percent complementarity" refers to the percentage of nucleobases of an oligomeric compound that are complementary to an equal-length portion of a target nucleic acid. Percent complementarity is calculated by dividing the total length of the oligomeric compound by the number of nucleobases in the oligomeric compound that are complementary to the nucleobases at the corresponding positions in the target nucleic acid.

As used herein, "percent identity" refers to a first nucleic acid that is of the same type (independent of chemical modification) as the nucleobase in the corresponding stomach in the second nucleic acid, divided by the total number of nucleobases in the first nucleic acid. means the number of nucleobases in

As used herein, “modulation” refers to a change in the amount or quality of a molecule, function, or activity as compared to the amount or quality of the molecule, function, or activity prior to modulation. For example, modulation includes a change, increase (stimulation or induction) or reduction (inhibition or reduction) in gene expression. As a further example, modulation of expression may include a change in splice site selection of pre-mRNA processing, wherein the absolute or relative amount of a particular splice-variant is changed compared to the amount in the absence of regulation.

As used herein, “chemical motif” refers to a pattern of chemical modifications in an oligonucleotide or region thereof. Motifs may be defined by modifications at specific nucleosides and/or specific linking groups of oligonucleotides.

As used herein, “nucleoside motif” refers to a pattern of nucleoside modifications in an oligonucleotide or region thereof. Linkages of such oligonucleotides may be modified or unmodified. Unless otherwise indicated, motifs herein that only describe nucleosides are intended to be nucleoside motifs. Accordingly, in such a case, the connection is not limited.

As used herein, “sugar motif” refers to a pattern of sugar modifications in an oligonucleotide or region thereof.

As used herein, “linkage motif” refers to a pattern of linkage modifications in an oligonucleotide or region thereof. The nucleosides of such oligonucleotides may be modified or unmodified. Unless otherwise indicated, motifs herein that merely describe linkages are intended to be linkage motifs. Thus, in such cases, the nucleosides are not limited.

As used herein, "nucleobase modification motif" refers to a pattern of modifications to a nucleobase along an oligonucleotide. Unless otherwise indicated, nucleobase modification motifs are independent of nucleobase sequence.

As used herein, “sequence motif” refers to a pattern of nucleobases arranged along an oligonucleotide or portion thereof. Unless otherwise indicated, sequence motifs are independent of chemical modifications and thus may have any combination of chemical modifications without including chemical modifications.

As used herein, "type of modification" in reference to a nucleoside or nucleoside of a "type" refers to a chemical modification of a nucleoside and includes modified and unmodified nucleosides. Thus, unless otherwise indicated, a “nucleoside having a modification of the first type” may be an unmodified nucleoside.

As used herein, "differently modified" refers to a chemical modification or chemical substitution that differs from one another, including the absence of modification. Thus, for example, MOE nucleosides and unmodified DNA nucleosides are “differently modified” even though the DNA nucleosides are unmodified. Likewise, DNA and RNA are "differently modified" even though both are naturally occurring unmodified nucleosides. Nucleosides comprising the same but different nucleobases are not modified differently. For example, a nucleoside comprising a 2'-OMe modified sugar and an unmodified adenine nucleobase and a nucleoside comprising a 2'-OMe modified sugar and an unmodified thymine nucleobase are not differently modified .

As used herein, "modifications of the same type" mean variations that are identical to each other, including the absence of variations. Thus, for example, two unmodified DNA nucleosides have "the same type of modification" even though the DNA nucleosides are unmodified. Such nucleosides with the same type modification may comprise different nucleobases.

As used herein, "distinct region" means a portion of an oligonucleotide, wherein the motif of the chemical modification or chemical modification of any contiguous division comprises at least one difference that allows the distinct regions to be distinguished from each other. do.

As used herein, "pharmaceutically acceptable carrier or diluent" means any substance suitable for use in administration to an animal. In certain embodiments, the pharmaceutically acceptable carrier or diluent is sterile saline. In certain embodiments, such sterile saline is pharmaceutical grade saline.

As used herein, the term “metabolic disorder” refers to a disease or condition characterized primarily by dysregulation of metabolism—a complex set of chemical reactions associated with the breakdown of food to produce energy.

As used herein, the term “cardiovascular disease” or “cardiovascular disorder” refers to a disease or condition characterized primarily by impaired function of the heart or blood vessels. Examples of cardiovascular diseases or disorders include, but are not limited to, aneurysms, angina pectoris, arrhythmias, atherosclerosis, cerebrovascular disease (stroke), coronary heart disease, hypertension, dyslipidemia, hyperlipidemia, and hypercholesterolemia.

The term "monocyclic or polycyclic ring system" as used herein is meant to include all ring systems selected from single or polycyclic radical ring systems, said rings being fused or linked, aliphatic, cycloaliphatic, aryl, heteroaryl, is meant to include single and mixed ring systems individually selected from aralkyl, arylalkyl, heterocyclic, heteroaryl, heteroaromatic and heteroarylalkyl. Such monocyclic or polycyclic cyclic structures contain rings each having the same level of saturation or each having, independently, varying degrees of saturation, including fully saturated, partially saturated or fully unsaturated. can do. Each ring may contain ring atoms selected from C, N, O and S to yield heterocyclic rings as well as rings containing only C ring atoms that may be present in mixed motifs such as benzimidazole, wherein one ring is unique It has carbon ring atoms and the fused ring has two nitrogen atoms. Monocyclic or polycyclic ring systems may be further substituted with substituent groups such as phthalimides having a 2 =O group attached to one of the rings. Monocyclic or polycyclic ring systems may be attached to the parent molecule using a variety of strategies, such as directly through ring atoms, and fused through multiple ring atoms, through substituent groups or through bifunctional linking moieties.

As used herein, "prodrug" refers to a compound in an inactive sub-active form, which, when administered to a subject, is metabolized to become active, or more active, a compound (eg, a drug) to form

As used herein, “substituent” and “substituent group” refer to an atom or group that replaces an atom or group of a named parent compound. For example, a substituent on a modified nucleoside is any atom or group that is different from the atom or group found in a naturally occurring nucleoside (e.g., a modified 2'-substituent is a nucleoside other than H or OH any atom or group in the 2'-position of Substituent groups can be protected or unprotected. In certain embodiments, compounds of the present disclosure have a substituent at one or more than one position of the parent compound. Substituents may also be further substituted with other substituent groups and may be attached to the parent compound either directly or through a linking group such as an alkyl or hydrocarbyl group.

Likewise, as used herein, “substituent” with respect to a chemical functional group means an atom or group of atoms that is different from the atom or group of atoms normally present in the named functional group. In certain embodiments, a substituent replaces a hydrogen atom of a functional group (e.g., in certain embodiments, a substituent on a substituted methyl group is an atom or group other than hydrogen that replaces one of the hydrogen atoms of an unsubstituted methyl group to be). Unless otherwise indicated, well-accepted groups as substituents include, but are not limited to, halogen, hydroxyl, alkyl, alkenyl, alkynyl, acyl (-C-(O)-R _aa ), carboxyl (- C(O)OR _aa ), aliphatic group, cycloaliphatic group, alkoxy, substituted oxy (-OR _aa ), aryl, aralkyl, heterocyclic radical, heteroaryl, heteroarylalkyl, amino (-N(R _bb ) -(R _cc )), imino(=NR _bb ), amido (-C(O)N-(R _bb )(R _cc ) or -N(R _bb )C(O)R _aa ), azido (-N ₃ ), nitro (-NO ₂ ), cyano (-CN), carbamido (-OC(O)N(R _bb )(R _cc ) or -N(R _bb )-C(O)- OR _aa ), ureido (-N(R _bb )C(O)-N(R _bb )(R _{cc )), thioureido} (-N(R bb )C-(S)N(R _bb ) -(R _cc )), guanidinyl (-N(R _bb )-C(=NR _bb )-N(R _bb )(R _cc )), amidinyl (-C(=NR _bb )N(R) _bb )(R _cc ) or -N(R _bb )C(=NR _bb )(R _aa )), thiol (-SR _bb ), sulfinyl (-S(O)R _bb ), sulfonyl (-S( O) ₂ R _bb ) and sulfonamidyl (-S(O) ₂ N(R _bb )(R _cc ) or -N(R _bb )-S-(O) ₂ R _bb . wherein each R _aa , R _bb and R _cc is, independently, H, an optionally linked chemical functional group or a further substituent group, a preferred list includes, but is not limited to, alkyl, alkenyl, alkynyl, aliphatic, alkoxy, acyl, aryl , aralkyl, heteroaryl, cycloaliphatic, heterocyclic and heteroarylalkyl. Selected substituents in the compounds described herein are present to a recurring degree.

As used herein, “alkyl,” as used herein, refers to a saturated straight-chain or branched hydrocarbon radical containing up to 24 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, isopropyl, n-hexyl, octyl, decyl, dodecyl, and the like. Alkyl groups typically contain from 1 to about 24 carbon atoms, more typically from 1 to about 12 carbon atoms (C ₁ -C ₁₂ alkyl), with 1 to about 6 carbon atoms being more preferred.

As used herein, "alkenyl" means a straight or branched hydrocarbon chain radical containing up to 24 carbon atoms and having at least one carbon-carbon double bond. Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, dienes such as 1,3-butadiene and the like. Alkenyl groups typically contain from 2 to about 24 carbon atoms, more typically from 2 to about 12 carbon atoms, with 2 to about 6 carbon atoms being more preferred. An alkenyl group, as used herein, may optionally include one or more additional substituent groups.

As used herein, “alkynyl” refers to a straight-chain or branched hydrocarbon radical containing up to 24 carbon atoms and having at least one carbon-carbon triple bond. Examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 1-butynyl, and the like. Alkynyl groups typically contain from 2 to about 24 carbon atoms, more typically from 2 to about 12 carbon atoms, with 2 to about 6 carbon atoms being more preferred. An alkynyl group, as used herein, may optionally include one or more additional substituent groups.

As used herein, "acyl" means a radical that will be formed by removal of a hydroxyl group from an organic acid and has the general formula -C(O)-X, where X is typically aliphatic, cycloaliphatic or aromatic. has Examples include aliphatic carbonyl, aromatic carbonyl, aliphatic sulfonyl, aromatic sulfinyl, aliphatic sulfinyl, aromatic phosphate, aliphatic phosphate, and the like. An acyl group, as used herein, may optionally include additional substituent groups.

As used herein, “cycloaliphatic” refers to a cyclic ring system, wherein the ring is aliphatic. A ring system may include one or more rings, wherein at least one ring is aliphatic. Preferred cycloaliphatic include rings having from about 5 to about 9 carbon atoms in the ring. Cycloaliphatic, as used herein, may optionally include additional substituent groups.

As used herein, "aliphatic" means a straight-chain or branched hydrocarbon radical containing up to 24 carbon atoms, wherein the saturation between any two carbon atoms is a single, double or triple bond. Aliphatic groups preferably contain from 1 to about 24 carbon atoms, more typically from 1 to about 12 carbon atoms, with 1 to about 6 carbon atoms being more preferred. The straight or branched chain of an aliphatic group may be interrupted with one or more heteroatoms including nitrogen, oxygen, sulfur and phosphorus. Such aliphatic groups interrupted by heteroatoms include, but are not limited to, polyalkoxys such as polyalkylene glycols, polyamines, and polyimines. Aliphatic groups, as used herein, may optionally include additional substituent groups.

As used herein, "alkoxy" refers to a radical formed between an alkyl group and an oxygen atom, which oxygen atom is used to attach the alkoxy group to the parent molecule. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, neopentoxy , n-hexoxy et al. Alkoxy groups, as used herein, may optionally include additional substituent groups.

As used herein, “aminoalkyl” refers to an amino substituted C ₁ -C ₁₂ alkyl radical. The alkyl portion of the radical forms a covalent bond with the parent molecule. The amino group may be at any position and the aminoalkyl group may be substituted with additional substituent groups at the alkyl and/or amino moieties.

As used herein, “aralkyl” and “arylalkyl” refer to an aromatic group covalently bonded to a _{C 1} -C _{12 alkyl radical.} The alkyl radical moiety of the resulting aralkyl (or arylalkyl) group forms a covalent bond with the parent molecule. Examples include, without limitation, benzyl, phenethyl, and the like. Aralkyl groups, as used herein, optionally include additional substituent groups attached to the alkyl, aryl, or both groups forming a radical group.

As used herein, “aryl” and “aromatic” refer to a monocyclic or polycyclic carbocyclic ring radical having one or more aromatic rings. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, idenyl, and the like. Preferred aryl ring systems have from about 5 to about 20 carbon atoms in one or more rings. Aryl groups, as used herein, may optionally include additional substituent groups.

As used herein, “halo” and “halogen” refer to an atom selected from fluorine, chlorine, bromine and iodine.

As used herein, “heteroaryl,” and “heteroaromatic” refer to a radical comprising a mono- or multi-cyclic aromatic ring, ring system or fused ring system, wherein at least one of the rings is aromatic and at least one contains heteroatoms. Heteroaryl is also meant to include fused ring systems, including systems in which at least one of the fused rings contains no heteroatoms. Heteroaryl groups typically contain one or more ring atoms selected from sulfur, nitrogen or oxygen. Examples of heteroaryl groups include, but are not limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isoxazolyl, thiadiazolyl, oxa diazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzooxazolyl, quinoxalinyl and the like. The heteroaryl radical may be attached to the parent molecule through a linking moiety such as an aliphatic group or a hetero atom. Heteroaryl groups, as used herein, may optionally include additional substituent groups.

As used herein, "conjugate compound" means any atom, group of atoms, or group of linked atoms suitable for use as a conjugate group. In certain embodiments, a conjugate compound may possess or confer one or more properties including, but not limited to, pharmacokinetic, pharmacodynamic, binding, uptake, cellular distribution, cellular uptake, charge and/or clearance properties. .

As used herein, unless otherwise indicated or modified, the term “double-stranded” refers to two distinct oligomeric compounds that are hybridized to one another. Such double-stranded compounds may have one or more or non-hybridizing nucleosides and/or one or more internal non-hybridizing nucleosides (mismatches) at one or both ends of one or both strands (overhangs). provided that there is sufficient complementarity to maintain hybridization under physiologically relevant conditions.

As used herein, “2′-O-methoxyethyl” (also 2′-MOE and 2′-O(CH ₂ ) ₂ -OCH ₃ ) refers to the O-methoxy- at the 2′ position of the furosyl ring ethyl modification. A 2'-0-methoxyethyl modified sugar is a modified sugar.

As used herein, “2′-O-methoxyethyl nucleotide” refers to a nucleotide comprising a 2′-O-methoxyethyl modified sugar moiety.

"3' target site" or "3' stop site" refers to a nucleotide of a target nucleic acid that is complementary to the 3'-most nucleotide of a particular antisense compound.

As used herein, "5' target site" or "5 initiation site" refers to a nucleotide of a target nucleic acid that is complementary to the 5'-most nucleotide of a particular antisense compound.

As used herein, "5-methylcytosine" refers to a cytosine modified by a methyl group attached to the 5' position. 5-Methylcytosine is a modified nucleobase.

As used herein, “about” means within ±10% of a value. For example, if it is stated that "the marker may be increased by about 50%", this suggests that the marker may increase between 45%-55%.

As used herein, "active pharmaceutical agent" means a substance or substances in a pharmaceutical composition that provides a therapeutic benefit when administered to a subject. For example, in certain embodiments, an antisense oligonucleotide targeted to ANGPTL3 is an active pharmaceutical agent.

As used herein, “active target region” or “target region” refers to a region to which one or more active antisense compounds are targeted.

As used herein, “active antisense compound” refers to an antisense compound that reduces target nucleic acid levels or protein levels.

As used herein, “adipogenesis” refers to the development of adipocytes from preadipocytes. "Lipogenesis" means the production or formation of fat, or fat degeneration or fat infiltration.

As used herein, "excess fat" or "obesity" refers to the condition of being obese, or an excessively large amount of body fat or adipose tissue in relation to lean body mass. The amount of body fat involves concern with both the distribution of fat throughout the body and the size and mass of adipose tissue deposits. Body fat distribution can be estimated by a skin-fold scale, waist-hip ratio, or techniques such as ultrasound, computed tomography, or magnetic resonance imaging. According to the Center for Disease Control and Prevention, individuals with a body mass index (BMI) of 30 or higher are considered obese. The term “obesity” as used herein includes a condition in which there is an increase in body fat that exceeds physical necessity as a result of excessive accumulation of adipose tissue in the body. The term “obesity” includes, but is not limited to, the following conditions: adult-type obesity; nutritional obesity; endogenous or metabolic obesity; endocrine obesity; familial obesity; high insulin obesity; hyperplastic-hypertrophic obesity; hypogonadism obesity; hypothyroidism obesity; Obesity throughout life; morbid obesity and extrinsic obesity.

As used herein, “administered simultaneously” refers to the co-administration of two agents in any manner in which the pharmacological effects of both are simultaneously exhibited in the patient. Concomitant administration does not require that both agents be administered in a single pharmaceutical composition, in the same dosage form, or by the same route of administration. The effects of both agents need not appear on their own at the same time. Effects only need to overlap for a period of time and do not need to span the same time (space).

As used herein, “administering” means providing an agent to an animal and includes, but is not limited to, administration by a healthcare professional and self-administration.

As used herein, "agent" refers to an active substance capable of providing a therapeutic benefit when administered to a mammal. By “first agent” is meant a therapeutic compound of the invention. For example, the first agent can be an antisense oligonucleotide that targets ANGPTL3. "Second agent" means a second therapeutic compound of the invention (eg, a second antisense oligonucleotide targeting ANGPTL3) and/or a non-ANGPTL3 therapeutic compound.

As used herein, "amelioration" refers to attenuating at least one indicator, sign, or symptom of an associated disease, disorder or condition. The severity of the marker can be determined by subjective or objective methods known to those skilled in the art.

"ANGPTL3" means any nucleic acid or protein of ANGPTL3.

"ANGPTL3 expression" means the level of mRNA transcribed from the gene encoding ANGPTL3 or the level of protein translated from mRNA. ANGPTL3 expression can be measured by methods known in the art, such as Northern or Western blot.

As used herein, “ANGPTL3 nucleic acid” refers to any nucleic acid encoding ANGPTL3. For example, in certain embodiments, an ANGPTL3 nucleic acid comprises a DNA sequence encoding ANGPTL3, a DNA encoding ANGPTL3 (introns and exons). RNA sequence transcribed from (including genomic DNA comprising

As used herein, "animal" means a human or non-human animal, which includes, but is not limited to, mice, rats, rabbits, dogs, cats, pigs, and monkeys and chimpanzees. non-human primates that have

As used herein, "apoB-containing lipoprotein" means any lipoprotein having apolipoprotein B as its protein component, and is intended to include LDL, VLDL, IDL, and lipoprotein (a). understood, and can generally be targeted by lipid lowering agents and therapies "ApoB-100-containing LDL" means the ApoB-100 isoform containing LDL.

As used herein, “atherosclerosis” means that hardening of arteries affecting large and medium-sized arteries is characterized by the presence of fat accumulation. Fat deposits are called "atheromas" or "plaques," which are composed primarily of cholesterol and other fats, calcium and scar tissue, and damage the linings of arteries.

As used herein, "cardiometabolic disease" or "cardiometabolic disorder" is a disease or disorder of both the cardiovascular and metabolic systems. Cardio-metabolic diseases or disorders include, but are not limited to, diabetes and dyslipidemia.

As used herein, “co-administration” refers to the administration of two or more agents to an individual. The two or more agents may be in a single pharmaceutical composition, or may be in separate pharmaceutical compositions. Each of the two or more agents may be administered via the same or different routes of administration Co-administration includes concurrent or sequential administration.

As used herein, "cholesterol" is a sterol molecule found in the cell membranes of all animal tissues. Cholesterol must be transported into the plasma of animals by lipoproteins, including very low density lipoproteins (VLDL), medium density lipoproteins (IDL), low density lipoproteins (LDL), and high density lipoproteins (HDL). "Plasma cholesterol" refers to the sum of all lipoprotein (VDL, IDL, LDL, HDL) esterified and/or non-estrified cholesterol present in plasma or serum.

As used herein, a “cholesterol absorption inhibitor” is an agent that inhibits the absorption of exogenous cholesterol obtained from a diet.

As used herein, "coronary heart disease (CHD)" refers to the narrowing of the small blood vessels supplying the heart with blood and oxygen, often as a result of atherosclerosis.

As used herein, "diabetes mellitus" or "diabetes" means a syndrome characterized by abnormal hyperglycemia (hyperglycemia) resulting from disordered metabolism and insufficient levels of insulin or reduced insulin sensitivity. Characteristic symptoms include excessive urine production due to high blood sugar levels (polyuria), excessive thirst to compensate for increased urination and increased fluid intake (polydipsia), blurred vision due to hyperglycemic effects on the optics of the eye, weight loss of unknown cause, and helplessness.

As used herein, "diabetic dyslipidemia" or "type 2 diabetes with dyslipidemia" refers to type 2 diabetes, decreased HDL-C, elevated triglycerides, and elevated small, dense LDL particles. a condition characterized by

As used herein, "diluent" refers to an ingredient in a composition that has no pharmacological activity but is pharmaceutically necessary or desirable. For example, the diluent in the injected composition may be a liquid, such as a saline solution.

As used herein, "dyslipidemia" refers to disorders of lipid and/or lipoprotein metabolism, including overproduction or deficiency of lipids and/or lipoproteins. Dyslipidemia can be manifested by elevations of cholesterol and triglycerides as well as lipoproteins such as low-density lipoprotein (LDL) cholesterol.

As used herein, "dosage unit" means the form in which the medicament is provided, eg, a pill, tablet, or other dosage unit known in the art. In certain embodiments, the dosage unit is a vial containing lyophilized antisense oligonucleotides. In certain embodiments, the dosage unit is a vial containing the reconstituted antisense oligonucleotide.

As used herein, “dose” refers to a specified amount of an agent given in a single administration or within a specified period of time. In certain embodiments, the dosage may be administered in one, two, or more boluses, tablets, or injections. For example, if subcutaneous administration is desired in certain embodiments, the desired dosage requires a volume not readily accommodated by a single injection, and thus, two or more injections may be used to achieve the desired dosage. In certain embodiments, the medicament is administered by infusion over an extended period of time or continuously. Dosage may be stated as an amount of drug per hour, day, week, or month. Dosage may be expressed in mg/kg or g/kg.

As used herein, "effective amount" or "therapeutically effective amount" means an amount of an active agent sufficient to cause a desired physiological result in a subject in need thereof. An effective amount may vary between individuals depending on the health and physical condition of the individual being treated, the taxon of the individual being treated, the formulation of the composition, assessment of the individual's medical condition, and other relevant factors.

As used herein, "glucose" is a monosaccharide used by cells as a source of energy and a metabolic intermediate. "Plasma glucose" means glucose present in plasma.

As used herein, “high-density lipoprotein-C” or “HDL-C” refers to cholesterol associated with high-density lipoprotein particles. The concentration of HDL-C in serum (or plasma) is typically quantified in mg/dL or nmol/L. "Serum HDL-C" and "plasma HDL-C" refer to HDL-C in serum and plasma, respectively.

As used herein, “HMG-CoA reductase inhibitor” refers to agents that act through inhibition of the enzyme HMG-CoA reductase, such as atorvastatin, rosuvastatin, fluvastatin, lovastatin, pravastatin, and simvastatin. .

As used herein, "hypercholesterolemia" means a condition characterized by elevated cholesterol or circulating (plasma) cholesterol, LDL-cholesterol and VLDL-cholesterol, according to the following guidelines: the Expert Panel Report of the National Cholesterol Educational Program (NCEP) of Detection, Evaluation of Treatment of high cholesterol in adults (see Arch. Int. Med. (1988) 148, 36-39).

As used herein, "hyperlipidemia" or "hyperlipidemia" is a condition characterized by elevated serum lipids or circulating (plasma) lipids. This condition represents an abnormally high concentration of fat. The lipid fractions in the circulating blood are cholesterol, low-density lipoproteins, ultra-low-density lipoproteins and triglycerides.

As used herein, “ultrahypertriglyceridemia” refers to a condition characterized by elevated triglyceride levels.

As used herein, “identifying” or “selecting a subject having a metabolic or cardiovascular disease” refers to identifying or selecting a subject diagnosed with a metabolic disease, cardiovascular disease, or metabolic disease, or a metabolic disease, cardiovascular disease, or metabolic disease. disease, or metabolic syndrome, such as, but not limited to, hypercholesterolemia, hyperglycemia, hyperlipidemia, hypertriglyceridemia, hypertension, increased insulin resistance, decreased insulin sensitivity, above normal weight, and/or above normal body fat content or specific thereof means identifying or selecting a subject having any syndrome of the combination. Such identification includes standard clinical tests or evaluations, such as measurement of serum or circulating (plasma) blood-glucose, measurement of serum or circulating (plasma) triglycerides, measurement of blood pressure, measurement of body fat content, measurement of body weight, etc. , but not limited thereto) may be achieved by any particular method.

As used herein, "identifying" or "selecting a diabetic subject" refers to identifying or selecting a subject identified as diabetic, or, such as, but not limited to, on an empty stomach of at least 110 mg/dL. means identifying or selecting a subject with any symptom of diabetes (type 1 or 2) with blood sugar, diabetes, polyuria, polydipsia, increased insulin resistance, and/or decreased insulin sensitivity.

As used herein, "identifying" or "selecting an obese subject" means identifying or selecting a subject identified as obese, or having a BMI greater than 30, and/or greater than 102 cm in males. means identifying or selecting a subject having a waist circumference, or a waist circumference greater than 88 cm in women.

As used herein, "identifying" or "selecting a dyslipidemic subject" refers to a subject identified with a lipid and/or lipoprotein metabolic disorder (including lipid and/or lipoprotein overproduction or deficiency). including identifying or selecting Dyslipidemia can be manifested by elevations of cholesterol and triglycerides as well as lipoproteins such as low-density lipoprotein (LDL) cholesterol.

As used herein, "identifying" or "selecting" a subject with increased adipose tissue includes concerns about the distribution of fat throughout the body and the size and mass of adipose tissue deposits ( or identifying or selecting a subject with an increased amount of adipose tissue). Body fat distribution can be estimated by a skin-fold scale, waist-hip ratio, or techniques such as ultrasound, computed tomography, or magnetic resonance imaging. According to the Center for Disease Control and Prevention, individuals with a body mass index (BMI) of 30 or higher are considered obese.

As used herein, “improved cardiovascular outcome” refers to the occurrence of, or a reduction in the risk of, adverse cardiovascular events. Examples of adverse cardiovascular events include, but are not limited to, death, re-infarction, stroke, cardiac shock, pulmonary edema, cardiac arrest, and atrial dysrhythmia.

As used herein, “immediately adjacent” means that there are no intervening elements between immediately adjacent elements.

As used herein, “individual” or “subject” or “animal” refers to a human or non-human animal selected for treatment or therapy.

As used herein, “insulin resistance” is defined as a condition in which normal amounts of insulin are inadequate to produce a normal insulin response from cells, such as fat, muscle and/or liver cells. Insulin resistance in gut cells results in hydrolysis of stored triglycerides, which raises free fatty acids in plasma. Insulin resistance in muscle reduces glucose uptake, whereas insulin resistance in the liver reduces glucose storage, the effect of both contributing to elevation of blood glucose. High plasma levels of insulin and glucose due to insulin resistance often lead to metabolic syndrome and type 2 diabetes.

As used herein, "insulin sensitivity" is a measure of how an individual processes glucose effectively. Individuals with high insulin sensitivity efficiently process glucose, whereas individuals with low insulin sensitivity do not process glucose effectively.

As used herein, “intravenous administration” refers to administration into a vein.

As used herein, “lipid-lowering” means a decrease in one or more lipids in a subject. Lipid-lowering may occur over time with one or more doses.

As used herein, “lipid-lowering agent” refers to an agent, eg, an ANGPTL3-specific modulator, provided to a subject to achieve lipid lowering in the subject. For example, in certain embodiments, the lipid-lowering agent is apoB, apoC-III, total cholesterol, LDL-C, VLDL-C, IDL-C, non-HDL-C, triglycerides, small dense LDL in the subject. particles, and at least one of Lp(a).

As used herein, “lipid-lowering therapy” refers to a therapeutic regimen given to a subject to reduce one or more lipids in the subject. In certain embodiments, the lipid-lowering therapy is apoB, apoC-III, total cholesterol, LDL-C, VLDL-C, IDL-C, non-HDL-C, triglycerides, small dense LDL particles, and Lp in the subject. provided to reduce one or more of (a).

As used herein, “lipoproteins”, such as VLDL, LDL and HDL, refer to a group of proteins found in serum, plasma and lymph and are important for lipid transport. The chemical composition of each lipoprotein is different in that HDL has a higher ratio of protein to lipid, whereas VLDL has a lower ratio of protein to lipid.

As used herein, "low density lipoprotein-cholesterol (LDL-C)" refers to cholesterol carried in low density lipoprotein particles. The concentration of LDL-C in serum (or plasma) is typically quantified in mg/dL or nmol/L. "Serum LDL-C" and "plasma LDL-C" refer to LDL-C in serum and plasma, respectively.

As used herein, “major risk factor” refers to a factor that contributes to a high risk for a particular disease or condition. In certain embodiments, major risk factors for coronary heart disease include, but are not limited to, cigarette smoking, high blood pressure, low HDL-C, family history of coronary heart disease, age, and other factors disclosed herein.

As used herein, “metabolic disorder” or “metabolic disease” refers to a condition characterized by alteration or disturbance of metabolic function. "Metabolic" and "metabolism" are terms well known in the art and generally encompass the full scope of biochemical processes that occur in living organisms. Metabolic disorders include, but are not limited to, hyperglycemia, prediabetes, diabetes (types I and 2), obesity, insulin resistance, metabolic syndrome and dyslipidemia due to type 2 diabetes.

As used herein, “metabolic syndrome” refers to a condition characterized by a clustering of lipid and non-lipid cardiovascular risk factors of subject origin. In certain embodiments, the metabolic syndrome is identified by any three of the following factors: waist circumference greater than 102 cm for men or greater than 88 cm for women; serum triglycerides of at least 150 mg/dL; HDL-C of less than 40 mg/dL in men or 50 mg/dL in women; blood pressure of at least 130/85 mmHg; and a fasting blood glucose of at least 110 mg/dL. These determinants can be readily measured in clinical examples (JAMA, 2001, 285: 2486-2497).

As used herein, “mixed dyslipidemia” refers to a condition characterized by elevated cholesterol and elevated triglycerides.

As used herein, “MTP inhibitor” refers to an agent that inhibits the enzyme microsomal triglyceride transfer protein.

As used herein, “non-alcoholic fatty liver disease” or “NAFLD” refers to a condition characterized by fatty inflammation of the liver that is not due to excessive alcohol use (eg, alcohol consumption of 20 g/day or more). . In certain embodiments, NAFLD is associated with insulin resistance and metabolic syndrome. NAFLD encompasses a spectrum of diseases ranging from simple triglyceride accumulation in hepatocytes (liver steatosis) to hepatic steatosis with inflammation (steatohepatitis), fibrosis, and cirrhosis.

As used herein, “non-alcoholic steatohepatitis” (NASH) results from the progression of NAFLD beyond the deposition of triglycerides. A “second hit” that can lead to necrosis, inflammation and fibrosis is necessary for the evolution of NASH. Second-hit candidates can be grouped into broad categories: factors that cause oxidative stress and factors that promote the expression of pro-inflammatory cytokines. Elevated hepatic triglycerides cause increased oxidative stress in animal and human hepatocytes, indicating a potential causal and effect relationship between hepatic triglyceride accumulation, oxidative stress, and progression of hepatic steatosis to NASH ( Browning and Horton, J Clin Invest, 2004 , 114 , 147-152). Hypertriglyceridemia and hyperlipidemia can lead to triglyceride accumulation in peripheral tissues (Shimamura et al., Biochem). Biophys Res Commun, 2004 , 322 , 1080-1085).

As used herein, “nucleic acid” refers to a molecule composed of monomeric nucleotides. Nucleic acids include ribonucleic acid (RNA), deoxyribonucleic acid (DNA), single-stranded nucleic acid, double-stranded nucleic acid, small intervening ribonucleic acid (siRNA), and microRNA (miRNA). Nucleic acids may also include combinations of these elements in a single molecule.

As used herein, “parenteral administration” refers to administration by any means other than through the digestive tract. Parenteral administration includes topical administration, subcutaneous administration, intravenous administration, intramuscular administration, intraarterial administration, intraperitoneal administration. intracranial administration, or intracranial administration, such as intrathecal or intraventricular administration, administration can be continuous, chronic, short-term or intermittent.

As used herein, "agent" refers to a substance that provides a therapeutic benefit when administered to a subject. For example, in certain embodiments, an antisense oligonucleotide targeted to ANGPTL3 is a pharmaceutical agent.

As used herein, "pharmaceutical composition" means a mixture of substances suitable for administration to a subject. For example, a pharmaceutical composition may comprise one or more active agents and a sterile aqueous solution.

As used herein, "pharmaceutically acceptable carrier" refers to a medium or diluent that does not interfere with the structure or function of an oligonucleotide. Certain of these carriers enable pharmaceutical compositions to be formulated, for example, into tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and lozenges for oral ingestion by a subject. Certain such carriers allow the pharmaceutical composition to be formulated for injection or infusion. For example, the pharmaceutically acceptable carrier can be a sterile aqueous solution.

As used herein, "pharmaceutically acceptable salts" are physiologically and pharmaceutically acceptable salts of antisense oligonucleotides, i.e., retain the desired biological activity of the parent compound and do not impart undesirable toxicological effects thereto. means no salt.

As used herein, “portion” refers to a defined number of contiguous (ie linked) nucleobases of a nucleic acid. In certain embodiments, a segment is a defined number of contiguous nucleobases of a target nucleic acid. In certain embodiments, a moiety refers to a defined number of contiguous nucleobases of an antisense compound.

As used herein, “preventing” means delaying or preventing the onset or development of a disease, disorder, or condition from a very small amount of time to an unspecified amount of time. Preventing also means reducing the risk of developing a disease, disorder, or condition.

As used herein, "adverse side effect" refers to a physiological response resulting from treatment in addition to the desired effect. In certain embodiments, side effects include injection site reactions, liver function test abnormalities, renal dysfunction, hepatotoxicity, renal toxicity, central nervous system abnormalities, myopathy, and malaise. For example, elevated levels of aminotransferase in serum may indicate hepatotoxicity or liver dysfunction. For example, increased bilirubin may indicate hepatotoxicity or liver dysfunction.

As used herein, "statin" refers to an agent that inhibits the activity of HMG-CoA reductase.

As used herein, “subcutaneous administration” refers to administration directly under the skin.

As used herein, “targeting” or “targeted” refers to the process of design and selection of an antisense compound that specifically hybridizes to a target nucleic acid and induces a desired effect.

As used herein, “target nucleic acid”, “target RNA”, and “target RNA transcript” all refer to a nucleic acid that can be targeted by an antisense compound.

As used herein, a “target region” is defined as a portion of a target nucleic acid having at least one identifiable structure, function, or characteristic.

As used herein, "target segment" refers to the sequence of nucleotides of a target nucleic acid to which one or more antisense compounds are targeted. "5' target site" or "5' initiation site" refers to the 5'-most nucleotide of the target segment. A “3′ target site” or “3′ stop site” refers to the 3′-most nucleotide of a target segment.

As used herein, “therapeutically effective amount” refers to an amount of an agent that provides a therapeutic benefit to an individual.

As used herein, “therapeutic lifestyle change” refers to dietary and lifestyle changes intended for low adipose/adipose tissue mass and/or cholesterol. These changes may reduce the risk of developing heart disease, as well as recommendations for dietary intake of total daily calories, total fat, saturated fat, polyunsaturated fat, monounsaturated fat, carbohydrates, protein, cholesterol, and insoluble fiber. Includes recommendations for physical activity.

As used herein, "triglyceride" refers to a lipid or neutral fat consisting of glycerol combined with three fatty acid molecules.

As used herein, "type 2 diabetes" (also known as "type 2 diabetes mellitus" or "diabetes mellitus, type 2", formerly known as "diabetes mellitus type 2", "non-insulin -dependent diabetes mellitus (NIDDM)", "obesity associated diabetes", or "adult-onset diabetes") is a metabolic disorder characterized primarily by insulin resistance, relative insulin deficiency, and hyperglycemia.

As used herein, “treat” refers to administration of a pharmaceutical composition to an animal to effect alteration or amelioration of a disease, disorder or condition.

specific embodiment

In certain embodiments, ANGPTL3 has the sequence set forth in Genbank Accession No. NM_014495.2 (incorporated herein as SEQ ID NO: 1). In certain embodiments, ANGPTL3 has the sequence set forth in Genbank Accession Number NT_032977.9 nucleotides 33032001 to 33046000 (herein incorporated as SEQ ID NO: 2).

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and is of SEQ ID NOs: 1-2 contain at least 8 contiguous nucleobases complementary to equal length portions.

In certain embodiments, the compounds comprise an siRNA or antisense oligonucleotide targeted to ANGPTL3 known in the art and a conjugate group described herein. Examples of antisense oligonucleotides targeted to ANGPTL3 suitable for conjugation include, but are not limited to, those disclosed in US 8,653,047 (WO 2011/085271), which is incorporated herein by reference in its entirety. In certain embodiments, the present compounds comprise an antisense oligonucleotide having the nucleobase sequence of any of SEQ ID NOs: 34-111 (disclosed in US 8,653,047) and a conjugate group. In certain embodiments, the compound comprises an siRNA sense or antisense strand (disclosed in US 8,653,047) having the nucleobase sequence of any of SEQ ID NOs: 34-111 and a conjugate group. The nucleobase sequences of all the aforementioned referenced SEQ ID NOs are incorporated herein by reference.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides in a length targeted to ANGPTL3. The ANGPTL3 target may have a sequence selected from any one of SEQ ID NOs: 1-2.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and nucleobase 1140 of SEQ ID NO: 1 to 1159 and a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases that are complementary to an equal length portion of 1159, wherein the nucleobase sequence of the modified oligonucleotide is at least 80% complementary to SEQ ID NO:1. In certain embodiments, the modified oligonucleotide is at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15 complementary to an equal length portion of nucleobases 1140 to 1159 of SEQ ID NO:1. , at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and nucleobase 1140 of SEQ ID NO: 1 to 1159, wherein the nucleobase sequence of the modified oligonucleotide is at least 80% complementary to SEQ ID NO:1.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and nucleobase 1907 of SEQ ID NO:1 to a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases complementary to an equal length portion of 1926, wherein the nucleobase sequence of the modified oligonucleotide is at least 80% complementary to SEQ ID NO:1. In certain embodiments, the modified oligonucleotide is at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15 complementary to an equal length portion of nucleobases 1907 to 1926 of SEQ ID NO: 1. , at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and nucleobase 1907 of SEQ ID NO:1 to 1926, wherein the nucleobase sequence of the modified oligonucleotide is at least 80% complementary to SEQ ID NO:1.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and nucleobase 147 of SEQ ID NO: 1 and a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases complementary to an equal length portion of to 162, wherein the nucleobase sequence of the modified oligonucleotide is at least 80% complementary to SEQ ID NO:1. In certain embodiments, the modified oligonucleotide is at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15 complementary to an equal length portion of nucleobases 147 to 162 of SEQ ID NO:1. , or at least 16 contiguous nucleobases.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and nucleobase 147 of SEQ ID NO: 1 to 162, wherein the nucleobase sequence of the modified oligonucleotide is at least 80% complementary to SEQ ID NO:1.

In certain embodiments, the modified oligonucleotide consists of 12-30, 15-30, 18-24, 19-22, 13-25, 14-25, 15-25 or 16-24 linked nucleosides. In certain embodiments, the modified oligonucleotide is 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 linked nucleosides or ranges defined by any two of these values. In certain embodiments, the length of the modified oligonucleotide is 16 linked nucleosides. In certain embodiments, the length of the modified oligonucleotide is 20 linked nucleosides.

at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15 modified oligonucleotides are complementary to equal length portions of SEQ ID NO: 1 or 2 , a nucleobase sequence comprising a portion of at least 16, at least 17, at least 18, at least 19, or at least 20 contiguous nucleobases.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and is selected from any of the following SEQ ID NOs: at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least has a nucleobase sequence comprising 19, or 20 contiguous nucleobases: 15-27, 30-73, 75-85, 87-232, 238, 240-243, 245-247, 249-262, 264-397 , 399-469, 471-541, 543-600, 604-760, 762-819, 821-966, 968-971, 973-975, 977-990, 992-1110, 1112-1186, 1188-1216, 1218 -1226, 1228-1279, 1281-1293, 1295-1304, 1306-1943, 1945-1951, 1953-1977, 1979-1981, 1983-2044, 2046-2097, 2099-2181, 2183-2232, 2234-2238 , 2240-2258, 2260-2265, 2267-2971, 2973-2976, 2978-4162, 4164-4329, 4331-4389, 4391-4394, 4396-4877.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and the nucleobase sequence of SEQ ID NO: 77 a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of have In certain embodiments, the compound comprises ISIS 563580 and a conjugate group. In certain embodiments, the compound consists of ISIS 563580 and a conjugate group.

In certain embodiments, the present disclosure provides a conjugated antisense compound represented by the structure: In certain embodiments, the antisense compound comprises a modified oligonucleotide ISIS 563580 having a 5'-X, wherein X is a conjugate group comprising GalNAc. In certain embodiments, the antisense compound consists of a modified oligonucleotide ISIS 563580 having a 5'-X, wherein X is a conjugate group comprising GalNAc.

In certain embodiments, the present disclosure provides a conjugated antisense compound represented by the structure: In certain embodiments, the antisense compound comprises a conjugated modified oligonucleotide ISIS 703801. In certain embodiments, the antisense compound consists of a conjugated modified oligonucleotide ISIS 703801.

In certain embodiments, the present disclosure provides a conjugated antisense compound represented by the structure: In certain embodiments, the antisense compound comprises a conjugated modified oligonucleotide ISIS 703802. In certain embodiments, the antisense compound consists of a conjugated modified oligonucleotide ISIS 703802.

In certain embodiments, the present disclosure provides a conjugated antisense compound represented by the structure: In certain embodiments, the antisense compound comprises a modified oligonucleotide having the nucleobase sequence of SEQ ID NO: 77, which has 5'-GalNAc with variability in the sugar modifications of the wing. In certain embodiments, the antisense compound consists of a modified oligonucleotide having the nucleobase sequence of SEQ ID NO: 77, which has 5'-GalNAc with variability in the sugar modifications of the wing.

wherein R ¹ is —OCH ₂ CH ₂ OCH ₃ (MOE) and R ² is H; or R ¹ and R ² together form a bridge, wherein R ¹ is —O— and R ² is —CH ₂ —, —CH(CH ₃ )—, or —CH ₂ CH ₂ —, and R ¹ and R ² is directly linked, and the bridge thereby obtained is selected from: -O-CH ₂ -, -O-CH(CH ₃ )-, and -O-CH ₂ CH ₂ -;

^{and for each pair of R 3} and R ⁴ on the same ring, independently for each ring: either R ³ is selected from H and —OCH ₂ CH ₂ OCH ₃ ^{and R 4} is H; or R ³ and R ⁴ together form a bridge, wherein R ³ is —O—, and R ⁴ is CH ₂ —, —CH(CH ₃ )—, or —CH ₂ CH ₂ — and R ³ and R ⁴ is directly linked, and the bridge thereby obtained is selected from: -O-CH ₂ -, -O-CH(CH ₃ )-, and -O-CH ₂ CH ₂ -;

and R ⁵ is selected from H and —CH _{3 ;}

and Z is selected from ^{S -} and O ^-.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and the nucleobase sequence of SEQ ID NO: 20 a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of have In certain embodiments, the compound comprises ISIS 544199 and a conjugate group. In certain embodiments, the compound consists of ISIS 544199 and a conjugate group.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and the nucleobase sequence of SEQ ID NO: 35 a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of have In certain embodiments, the compound comprises ISIS 560400 and a conjugate group. In certain embodiments, the compound consists of ISIS 560400 and a conjugate group.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and the nucleobase sequence of SEQ ID NO: 90 a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of have In certain embodiments, the compound comprises ISIS 567233 and a conjugate group. In certain embodiments, the compound consists of ISIS 567233 and a conjugate group.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and the nucleobase sequence of SEQ ID NO: 93 a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of have In certain embodiments, the compound comprises ISIS 567320 and a conjugate group. In certain embodiments, the compound consists of ISIS 567320 and a conjugate group.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and the nucleobase sequence of SEQ ID NO: 94 a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or 20 contiguous nucleobases of have In certain embodiments, the compound comprises ISIS 567321 and a conjugate group. In certain embodiments, the compound consists of ISIS 567321 and a conjugate group.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and the nucleobase sequence of SEQ ID NO: 110 has a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, or 16 contiguous nucleobases of In certain embodiments, the compound comprises ISIS 559277 and a conjugate group. In certain embodiments, the compound consists of ISIS 559277 and a conjugate group.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides and the nucleobase sequence of SEQ ID NO: 114 has a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, or 16 contiguous nucleobases of In certain embodiments, the compound comprises ISIS 561011 and a conjugate group. In certain embodiments, the compound consists of ISIS 561011 and a conjugate group.

In certain embodiments, the nucleobase sequence of the modified oligonucleotide is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% complementary to SEQ ID NOs: 1-2. (as measured across the modified oligonucleotides).

In certain embodiments, the compounds disclosed herein are single-stranded oligonucleotides. In certain embodiments, the compounds disclosed herein are single-stranded modified oligonucleotides.

In certain embodiments, at least one internucleoside linkage of the modified oligonucleotide is a modified internucleoside linkage. In certain embodiments, the modified internucleoside linkage is a phosphorothioate internucleoside linkage. In certain embodiments, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 internucleoside linkages of the modified oligonucleotide are phosphorothio It is an eight internucleoside linkage. In certain embodiments, each internucleoside linkage is a phosphorothioate internucleoside linkage. In certain embodiments, the modified oligonucleotide comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9 or at least 10 phosphodiester internucleoside linkages do. In certain embodiments, each internucleoside linkage of the modified oligonucleotide is selected from a phosphodiester internucleoside linkage and a phosphorothioate internucleoside linkage.

In certain embodiments, at least one nucleoside of the modified oligonucleotide comprises a modified sugar. In certain embodiments, the at least one modified sugar is a bicyclic sugar. at least one modified sugar is 2'-0-methoxyethyl, restricted ethyl, 3'-fluoro-HNA or 4'-(CH ₂ ) _n -O-2' bridge, wherein n is 1 or a compound of 2.

In certain embodiments, at least one nucleoside of the modified oligonucleotide comprises a modified nucleobase. In certain embodiments, the modified nucleobase is 5-methylcytosine.

Certain embodiments disclosed herein provide a compound or composition comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide has: a) a gap segment consisting of linked deoxynucleosides; b) a 5' wing segment consisting of linked nucleosides; and c) a 3' wing segment consisting of linked nucleosides. The gap segment is located between the 5' wing segment and the 3' wing segment, and each nucleoside of each wing segment comprises a modified sugar.

In certain embodiments, the modified oligonucleotide consists of 12 to 30 linked nucleosides, comprising: a gap segment consisting of linked deoxynucleosides; a 5' wing segment consisting of linked nucleosides; a 3' wing segment consisting of linked nucleosides; wherein the gap segment is located between the 5' wing segment and the 3' wing segment, and each nucleoside of each wing segment comprises a modified sugar.

In certain embodiments, a compound or composition disclosed herein is a modified oligo consisting of 20 linked nucleosides having a nucleobase sequence comprising at least 8 contiguous nucleobases complementary to an equal length portion of SEQ ID NOs: 1-2. nucleotides, wherein the modified oligonucleotides comprise: a gap segment consisting of 10 linked deoxynucleosides; 5' wing segment consisting of 5 linked nucleosides; a 3' wing segment consisting of 5 linked nucleosides; wherein said gap segment is located between the 5' wing segment and the 3' wing segment, and each nucleoside of each wing segment comprises a 2'-0-methoxyethyl sugar, wherein at least one internucleoside The linkage is a phosphorothioate linkage, wherein each cytosine residue is 5-methylcytosine. In certain embodiments, each internucleoside linkage is a phosphorothioate linkage.

In certain embodiments, the modified oligonucleotide consists of 20 linked nucleosides and comprises: a gap segment of 10 linked deoxynucleosides; 5' wing segment consisting of 5 linked nucleosides; a 3' wing segment consisting of 5 linked nucleosides; wherein said gap segment is located between the 5' wing segment and the 3' wing segment, and each nucleoside of each wing segment comprises a 2'-0-methoxyethyl sugar, wherein at least one internucleoside The linkage is a phosphorothioate linkage, wherein each cytosine residue is 5-methylcytosine. In certain embodiments, each internucleoside linkage is a phosphorothioate linkage.

In certain embodiments, a compound or composition disclosed herein comprises a modified oligonucleotide consisting of 20 linked nucleosides having a nucleobase sequence comprising at least 8 contiguous nucleobases of the selected nucleobase sequence of SEQ ID NO: 77. and wherein the modified oligonucleotide comprises: a gap segment consisting of 10 linked deoxynucleosides; 5' wing segment consisting of 5 linked nucleosides; a 3' wing segment consisting of 5 linked nucleosides; wherein said gap segment is located between the 5' wing segment and the 3' wing segment, and each nucleoside of each wing segment comprises a 2'-0-methoxyethyl sugar, wherein at least one internucleoside The linkage is a phosphorothioate linkage, wherein each cytosine residue is 5-methylcytosine. In certain embodiments, each internucleoside linkage is a phosphorothioate linkage.

In certain embodiments, the modified oligonucleotide consists of 20 linked nucleosides having the nucleobase sequence of SEQ ID NO: 77, comprising: a gap segment consisting of 10 linked deoxynucleosides; 5' wing segment consisting of 5 linked nucleosides; a 3' wing segment consisting of 5 linked nucleosides; wherein said gap segment is located between the 5' wing segment and the 3' wing segment, and each nucleoside of each wing segment comprises a 2'-0-methoxyethyl sugar, wherein at least one internucleoside The linkage is a phosphorothioate linkage, wherein each cytosine residue is 5-methylcytosine. In certain embodiments, each internucleoside linkage is a phosphorothioate linkage.

In certain embodiments, a compound or composition disclosed herein comprises a modified oligonucleotide consisting of 20 linked nucleosides having a nucleobase sequence comprising at least 8 contiguous nucleobases of the selected nucleobase sequence of SEQ ID NO: 20 and wherein the modified oligonucleotide comprises: a gap segment consisting of 10 linked deoxynucleosides; 5' wing segment consisting of 5 linked nucleosides; a 3' wing segment consisting of 5 linked nucleosides; wherein said gap segment is located between the 5' wing segment and the 3' wing segment, and each nucleoside of each wing segment comprises a 2'-0-methoxyethyl sugar, wherein at least one internucleoside The linkage is a phosphorothioate linkage, wherein each cytosine residue is 5-methylcytosine. In certain embodiments, each internucleoside linkage is a phosphorothioate linkage.

In certain embodiments, the modified oligonucleotide consists of 20 linked nucleosides having the nucleobase sequence of SEQ ID NO: 20, comprising: a gap segment consisting of 10 linked deoxynucleosides; 5' wing segment consisting of 5 linked nucleosides; a 3' wing segment consisting of 5 linked nucleosides; wherein said gap segment is located between the 5' wing segment and the 3' wing segment, and each nucleoside of each wing segment comprises a 2'-0-methoxyethyl sugar, wherein at least one internucleoside The linkage is a phosphorothioate linkage, wherein each cytosine residue is 5-methylcytosine. In certain embodiments, each internucleoside linkage is a phosphorothioate linkage.

In certain embodiments, a compound or composition disclosed herein comprises a modified oligonucleotide consisting of 16 linked nucleosides having a nucleobase sequence comprising at least 8 contiguous nucleobases of the selected nucleobase sequence of SEQ ID NO: 110 and wherein the modified oligonucleotide comprises: a gap segment consisting of 10 linked deoxynucleosides; a 5' wing segment consisting of three linked nucleosides; a 3' wing segment consisting of three linked nucleosides; wherein said gap segment is located between the 5' wing segment and the 3' wing segment, each wing segment comprising a 2'-0-methoxyethyl sugar and at least one cEt sugar, wherein at least one internucleoside The linkage is a phosphorothioate linkage, wherein each cytosine residue is 5-methylcytosine. In certain embodiments, each internucleoside linkage is a phosphorothioate linkage.

In certain embodiments, the modified oligonucleotide consists of 16 linked nucleosides having the nucleobase sequence of SEQ ID NO: 110, comprising: a gap segment consisting of 10 linked deoxynucleosides; a 5' wing segment consisting of three linked nucleosides; a 3' wing segment consisting of three linked nucleosides; wherein said gap segment is located between the 5' wing segment and the 3' wing segment, each wing segment comprising a 2'-0-methoxyethyl sugar and at least one cEt sugar, wherein at least one internucleoside The linkage is a phosphorothioate linkage, wherein each cytosine residue is 5-methylcytosine. In certain embodiments, each internucleoside linkage is a phosphorothioate linkage.

In certain embodiments, the conjugate group is linked to the modified oligonucleotide at the 5' end of the modified oligonucleotide. In certain embodiments, the conjugate group is linked to the modified oligonucleotide at the 3' end of the modified oligonucleotide.

In certain embodiments, the conjugate group comprises exactly one ligand. In certain embodiments, the conjugate group comprises one or more ligands. In certain embodiments, the conjugate group comprises exactly two ligands. In certain embodiments, the conjugate group comprises two or more ligands. In certain embodiments, the conjugate group comprises three or more ligands. In certain embodiments, the conjugate group comprises exactly three ligands. In certain embodiments, each ligand is a compound selected from: polysaccharides, modified polysaccharides, mannose, galactose, mannose derivatives, galactose derivatives, D-mannopyranose, L-mannopyranose, D-arabinose Lose, L-galactose, D-xylofuranose, L-xylofuranose, D-glucose, L-glucose, D-galactose, L-galactose, α-D-mannofuranose, β-D-mannofura North, α-D-mannopyranose, β-D-mannopyranose, α-D-glucopyranose, β-D-glucopyranose, α-D-glucofuranose, β-D-glucofura North, α-D-fructofuranose, α-D-fructopyranose, α-D-galactopyranose, β-D-galactopyranose, α-D-galactofuranose, β-D -Galactofuranose, glucosamine, sialic acid, α-D-galactosamine, N-acetylgalactosamine, 2-amino-3- O -[( R )-1-carboxyethyl]-2-deoxy- β-D-glucopyranose, 2-deoxy-2-methylamino-L-glucopyranose, 4,6-dideoxy-4-formamido-2,3-di- O -methyl-D-only nopyranose, 2-deoxy-2-sulfoamino-D-glucopyranose, N -glycoloyl-α-neuraminic acid, 5-thio-β-D-glucopyranose, methyl 2,3, 4-tri- O -acetyl-1-thio-6- O -trityl-α-D-glucopyranoside, 4-thio-β-D-galactopyranose, ethyl 3,4,6,7- tetra- O -acetyl-2-deoxy-1,5-dithio-α-D- gluco -heptopyranoside, 2,5-anhydro-D-alononitrile, ribose, D-ribose, D- 4-thioribose, L-ribose, L-4-thioribose. In certain embodiments, each ligand is N-acetyl galactosamine.

In certain embodiments, the conjugate group comprises:

In certain embodiments, the conjugate group comprises:

In certain embodiments, the conjugate group comprises:

In certain embodiments, the conjugate group comprises:

In certain embodiments, the conjugate group comprises:

.

.

In certain embodiments, the conjugate group comprises at least one of a phosphorus linking group or a neutral linking group.

In certain embodiments, the conjugate group comprises a structure selected from:

;

;

wherein n is 1 to 12; and

m is 1 to 12.

In certain embodiments, the conjugate group has a linking group having a structure selected from:

wherein L is a phosphorus linking group or a neutral linking group;

Z ₁ is C(=O)OR ₂ ;

Z ₂ is H, C ₁ -C ₆ alkyl or substituted C ₁ -C ₆ alkynyl (alky), and;

R ₂ is H, C ₁ -C ₆ alkyl or substituted C ₁ -C ₆ alkynyl (alky), and; and

each m ₁ is, independently, 0 to 20, wherein at least one m ₁ is greater than 0 for each linking group.

In certain embodiments, the conjugate group has a linking group having a structure selected from:

wherein Z ₂ is H or CH ₃ ; and

each m ₁ is, independently, 0 to 20, wherein at least one m ₁ is greater than 0 for each linking group.

In certain embodiments, the conjugate group has a linking group having a structure selected from:

;

;

wherein n is 1 to 12; and

m is 1 to 12.

In certain embodiments, the conjugate group is covalently linked to the modified oligonucleotide.

In certain embodiments, the compound has a structure represented by the formula:

In the above formula,

A is a modified oligonucleotide;

B is a cleavable moiety;

C is a conjugate linker

D is a branching group

each E is a linking group;

each F is a ligand; and

q is an integer from 1 to 5;

In certain embodiments, the compound has a structure represented by the formula:

In the above formula,

A is a modified oligonucleotide;

B is a cleavable moiety;

C is a conjugate linker

D is a branching group

each E is a linking group;

each F is a ligand;

each n is independently 0 or 1; and

q is an integer from 1 to 5;

In certain embodiments, the compound has a structure represented by the formula:

In the above formula,

A is a modified oligonucleotide;

B is a cleavable moiety;

C is a conjugate linker

each E is a linking group;

each F is a ligand; and

q is an integer from 1 to 5;

In certain embodiments, the compound has a structure represented by the formula:

In the above formula,

A is a modified oligonucleotide;

C is a conjugate linker

D is a branching group;

each E is a linking group;

each F is a ligand; and

q is an integer from 1 to 5;

In certain embodiments, the compound has a structure represented by the formula:

In the above formula,

A is a modified oligonucleotide;

C is a conjugate linker

each E is a linking group;

each F is a ligand; and

q is an integer from 1 to 5;

In certain embodiments, the compound has a structure represented by the formula:

In the above formula,

A is a modified oligonucleotide;

B is a cleavable moiety;

D is a branching group;

each E is a linking group;

each F is a ligand; and

q is an integer from 1 to 5;

In certain embodiments, the compound has a structure represented by the formula:

In the above formula,

A is a modified oligonucleotide;

B is a cleavable moiety;

each E is a linking group;

each F is a ligand; and

q is an integer from 1 to 5;

In certain embodiments, the compound has a structure represented by the formula:

In the above formula,

A is a modified oligonucleotide;

D is a branching group;

each E is a linking group;

each F is a ligand; and

q is an integer from 1 to 5;

In certain embodiments, the conjugate linker has a structure selected from:

.

.

wherein each L is, independently, a phosphorus linking group or a neutral linking group; and

each n is, independently, 1-20.

In certain embodiments, the conjugate linker has a structure selected from:

.

.

In certain embodiments, the conjugate linker has the structure:

.

.

In certain embodiments, the conjugate linker has a structure selected from:

.

.

In certain embodiments, the conjugate linker has a structure selected from:

.

.

In certain embodiments, the conjugate linker has a structure selected from:

In certain embodiments, the conjugate linker comprises pyrrolidine. In certain embodiments, the conjugate linker does not comprise pyrrolidine.

In certain embodiments, the conjugate linker comprises PEG.

In certain embodiments, the conjugate linker comprises an amide. In certain embodiments, the conjugate linker comprises at least two amides. In certain embodiments, the conjugate linker does not comprise an amide. In certain embodiments, the conjugate linker comprises a polyamide.

In certain embodiments, the conjugate linker comprises an amine.

In certain embodiments, the conjugate linker comprises one or more disulfide bonds.

In certain embodiments, the conjugate linker comprises a protein binding moiety. In certain embodiments, the protein binding moiety comprises a lipid. In certain embodiments, the protein binding moiety is selected from cholesterol, cholic acid, adamantane acetic acid, 1-pyrene butyric acid, dihydrotestosterone, 1,3-bis-O(hexadecyl)glycerol, geranyloxy Hexyl group, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecyl group, palmitic acid, myristic acid, O3-(oleoyl) lithocholic acid, O3-(oleoyl) cholenic acid, dimethoxytrityl, or phenoxazine), vitamins (eg folate, vitamin A, vitamin E, biotin, pyridoxal), peptides, carbohydrates (eg monosaccharides, disaccharides, trisaccharides, tetras saccharides, oligosaccharides, polysaccharides), endosomal components, steroids (eg ubaol, hexigenin, diosgenin), terpenes (eg triterpenes, eg sarsapogenin, free delrin, epipriedelanol derived lithocholic acid), or cationic lipids. In certain embodiments, the protein binding moiety is selected from: C16 to C22 long chain saturated or unsaturated fatty acids, cholesterol, cholic acid, vitamin E, adamantane or 1-pentafluoropropyl.

In certain embodiments, the conjugate linker has a structure selected from:

wherein each n is independently 1 to 20; and p is 1 to 6.

In certain embodiments, the conjugate linker has a structure selected from:

In the above formula, each n is independently 1 to 20.

In certain embodiments, the conjugate linker has a structure selected from:

.

.

In certain embodiments, the conjugate linker has a structure selected from:

In the above formula, n is 1 to 20.

In certain embodiments, the conjugate linker has a structure selected from:

In certain embodiments, the conjugate linker has a structure selected from:

;

;

wherein each n is independently 0, 1, 2, 3, 4, 5, 6, or 7.

In certain embodiments, the conjugate linker has the structure:

.

.

In certain embodiments, the branching group has one of the following structures:

wherein each A1 is independently O, S, C=O or NH; and

each n is, independently, 1-20.

In certain embodiments, the branching group has one of the following structures:

wherein each A1 is independently O, S, C=O or NH; and

each n is, independently, 1-20.

In certain embodiments, the branching group has the structure:

.

.

In certain embodiments, the branching group has the structure:

.

.

In certain embodiments, the branching group has the structure:

.

.

In certain embodiments, the branching group has the structure:

.

.

In certain embodiments, the branching group comprises an ether.

In certain embodiments, the branching group has the structure:

wherein each n is independently 1 to 20; and

m is 2 to 6.

In certain embodiments, the branching group has the structure:

.

.

In certain embodiments, the branching group has the structure:

.

.

In certain embodiments, the branching group comprises:

;

;

wherein each j is an integer 1 to 3; and

each n is an integer from 1 to 20.

In certain embodiments, the branching group comprises:

.

.

In certain embodiments, each linking group is selected from:

wherein L is selected from a phosphorus linking group and a neutral linking group;

Z ₁ is C(=O)OR ₂ ;

Z ₂ is H, C ₁ -C ₆ alkyl or substituted C ₁ -C ₆ alkynyl (alky), and;

R ₂ is H, C ₁ -C ₆ alkyl or substituted C ₁ -C ₆ alkynyl (alky), and; and

each m ₁ is, independently, 0 to 20, wherein at least one m ₁ is greater than 0 for each linking group.

In certain embodiments, each linking group is selected from:

wherein Z ₂ is H or CH ₃ ; and

each m ₂ is, independently, 0 to 20, wherein at least one m ₂ is greater than 0 for each linking group.

In certain embodiments, each linking group is selected from:

;

;

wherein n is 1 to 12; and

m is 1 to 12.

In certain embodiments, at least one linking group comprises ethylene glycol.

In certain embodiments, at least one linking group comprises an amide. In certain embodiments, the at least one linking group comprises a polyamide.

In certain embodiments, at least one linking group comprises an amine.

In certain embodiments, at least two linking groups are different from each other. In certain embodiments, all linking groups are identical to each other.

In certain embodiments, each linking group is selected from:

wherein each n is, independently, 1 to 20; and

each p is 1 to about 6.

In certain embodiments, each linking group is selected from:

In certain embodiments, each linking group has the structure:

In the above formula, each n is independently 1 to 20.

In certain embodiments, each linking group has the structure:

.

.

In certain embodiments, the linking group has a structure selected from:

; 여기서 각각의 n은 독립적으로, 0, 1, 2, 3, 4, 5, 6, 또는 7이다.

; wherein each n is independently 0, 1, 2, 3, 4, 5, 6, or 7.

In certain embodiments, the linking group has a structure selected from:

.

.

In certain embodiments, the ligand is galactose.

In certain embodiments, the ligand is mannose-6-phosphate.

In certain embodiments, each ligand is selected from:

wherein each R ₁ is selected from OH and NHCOOH.

In certain embodiments, each ligand is selected from:

.

.

In certain embodiments, each ligand has the structure:

.

.

In certain embodiments, each ligand has the structure:

.

.

In certain embodiments, the conjugate group comprises a cell-targeting moiety.

In certain embodiments, the conjugate group comprises a cell-targeting moiety having the structure:

In the above formula, each n is independently 1 to 20.

In certain embodiments, the cell-targeting moiety has the structure:

.

.

In certain embodiments, the cell-targeting moiety has the structure:

In the above formula, each n is independently 1 to 20.

In certain embodiments, the cell-targeting moiety has the structure:

In certain embodiments, the cell-targeting moiety comprises:

.

.

In certain embodiments, the cell-targeting moiety comprises:

.

.

In certain embodiments, the cell-targeting moiety has the structure:

.

.

In certain embodiments, the cell-targeting moiety has the structure:

.

.

In certain embodiments, the cell-targeting moiety comprises:

.

.

In certain embodiments, the cell-targeting moiety has the structure:

.

.

In certain embodiments, the cell-targeting moiety comprises:

.

.

In certain embodiments, the cell-targeting moiety comprises:

.

.

In certain embodiments, the cell-targeting moiety comprises:

.

.

In certain embodiments, the cell-targeting moiety has the structure:

.

.

In certain embodiments, the cell-targeting moiety has the structure:

.

.

In certain embodiments, the cell-targeting moiety has the structure:

.

.

In certain embodiments, the cell-targeting moiety has the structure:

.

.

In certain embodiments, the cell-targeting moiety has the structure:

.

.

In certain embodiments, the cell-targeting moiety comprises:

.

.

In certain embodiments, the cell-targeting moiety comprises:

.

.

In certain embodiments, the cell-targeting moiety comprises:

.

.

In certain embodiments, the cell-targeting moiety comprises:

.

.

In certain embodiments, the cell-targeting moiety has the structure:

.

.

In certain embodiments, the cell-targeting moiety comprises:

.

.

In certain embodiments, the cell-targeting moiety has the structure:

.

.

In certain embodiments, the cell-targeting moiety comprises:

;

;

wherein each Y is selected from O, S, substituted or unsubstituted C ₁ -C ₁₀ alkyl, amino, substituted amino, azido, alkenyl or alkynyl.

In certain embodiments, the conjugate group comprises:

;

;

wherein each Y is selected from O, S, substituted or unsubstituted C ₁ -C ₁₀ alkyl, amino, substituted amino, azido, alkenyl or alkynyl.

In certain embodiments, the cell-targeting moiety has the structure:

;

;

wherein each Y is selected from O, S, substituted or unsubstituted C ₁ -C ₁₀ alkyl, amino, substituted amino, azido, alkenyl or alkynyl.

In certain embodiments, the conjugate group comprises:

In certain embodiments, the conjugate group comprises:

In certain embodiments, the conjugate group comprises:

In certain embodiments, the conjugate group comprises:

In certain embodiments, the conjugate group comprises a cleavable moiety selected from: a phosphodiester, an amide, or an ester.

In certain embodiments, the conjugate group comprises a phosphodiester cleavable moiety.

In certain embodiments, the conjugate group does not include a cleavable moiety, and the conjugate group includes a phosphorothioate linkage between the conjugate group and the oligonucleotide.

In certain embodiments, the conjugate group comprises an amide cleavable moiety.

In certain embodiments, the conjugate group comprises an ester cleavable moiety.

In certain embodiments, the compound has the structure:

In the above formula, each n is independently 1 to 20.

Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, the compound has the structure:

In the above formula, each n is independently 1 to 20.

Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, the compound has the structure:

In the above formula, each n is independently 1 to 20.

Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide;

Z is H or a linked solid support; and

Bx is a heterocyclic base moiety.

In certain embodiments, the compound has the structure:

In the above formula, each n is independently 1 to 20.

Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide;

Z is H or a linked solid support; and

Bx is a heterocyclic base moiety.

In certain embodiments, the compound has the structure:

wherein Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, the compound has the structure:

,

,

wherein Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, the compound has the structure:

,

,

wherein Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, the compound has the structure:

wherein Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, the compound has the structure:

wherein Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, the compound has the structure:

wherein Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, the compound has the structure:

wherein Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, the compound has the structure:

wherein Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, the compound has the structure:

wherein Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, the compound has the structure:

wherein Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, the compound has the structure:

wherein Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, the conjugate group comprises:

wherein Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, the conjugate group comprises:

wherein Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, the conjugate group comprises:

wherein Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ;

A is a modified oligonucleotide; and

Bx is a heterocyclic base moiety.

In certain embodiments, B _x is selected from adenine, guanine, thymine, uracil, or cytosine, or 5-methyl cytosine. In certain embodiments, B _x is adenine. In certain embodiments, B _x is thymine. Q ₁₃ is H or O(CH ₂ ) ₂ —OCH ₃ ; In certain embodiments, Q ₁₃ is H.

Certain embodiments of the present invention provide prodrugs comprising a composition or compound disclosed herein. Certain embodiments provide methods of using the conjugated antisense compounds and compositions described herein to inhibit ANGPTL3 expression. In certain embodiments, the conjugated antisense compound or composition comprises at least 5%, at least 10%, at least 20%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55% ANGPTL3. , at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95%. In a preferred embodiment, the antisense compound comprising a modified oligonucleotide and a conjugate group reduces ANGPTL3 by at least 50%. In a preferred embodiment, the antisense compound comprising a modified oligonucleotide and a conjugate group reduces ANGPTL3 by at least 55%. In a preferred embodiment, the antisense compound comprising a modified oligonucleotide and a conjugate group reduces ANGPTL3 by at least 60%. In a preferred embodiment, an antisense compound comprising a modified oligonucleotide and a conjugate group reduces ANGPTL3 by at least 65%. In a preferred embodiment, the antisense compound comprising a modified oligonucleotide and a conjugate group reduces ANGPTL3 by at least 70%. In a preferred embodiment, the antisense compound comprising a modified oligonucleotide and a conjugate group reduces ANGPTL3 by at least 75%. In a preferred embodiment, the antisense compound comprising a modified oligonucleotide and a conjugate group reduces ANGPTL3 by at least 80%. In a preferred embodiment, the antisense compound comprising a modified oligonucleotide and a conjugate group reduces ANGPTL3 by at least 85%. In a preferred embodiment, the antisense compound comprising a modified oligonucleotide and a conjugate group reduces ANGPTL3 by at least 90%. In a preferred embodiment, the antisense compound comprising a modified oligonucleotide and a conjugate group reduces ANGPTL3 by at least 95%.

_{In certain embodiments, a conjugated antisense compound or composition described herein has an in vitro IC 50} of less than 20 μM, less than 10 μM, less than 8 μM, less than 5 μM, less than 2 μM, less than 1 μM, or less than 0.8 μM (Example 2). in Hep3B cell lines as described in -3 and 7-10).

In certain embodiments, a conjugated antisense compound or composition described herein has a viscosity of less than 40 cP, less than 35 cP, less than 30 cP, less than 25 cP, as measured by a parameter (as described in Example 13). , less than 20 cP, or less than 15 cP.

In certain embodiments, the conjugated antisense compounds or compositions described herein are administered in vivo as described in the Examples. As evidenced by tolerance measurements, it is highly resistant. In certain embodiments, certain conjugated antisense compounds as described herein have an increase in ALT and/or AST values of no more than 4-fold, 3-fold, 2-fold, or 1.5-fold less than that of a saline treated animal. As evidenced by this, it is highly resistant.

Certain embodiments described herein provide salts of the conjugated antisense compounds disclosed herein. In certain embodiments, a compound or composition disclosed herein comprises a salt of a conjugate group and a modified oligonucleotide.

In certain embodiments, a conjugated antisense compound or composition disclosed herein further comprises a pharmaceutically acceptable carrier or diluent.

In certain embodiments, the animal is a human

Certain embodiments disclosed herein provide methods comprising administering to an animal a conjugated antisense compound or composition disclosed herein. In certain embodiments, the conjugated antisense compound or composition is therapeutic. In certain embodiments, administering a conjugated antisense compound or composition treats, prevents, or delays the progression of a disease associated with ANGPTL3. In certain embodiments, the disease relates to elevated ANGPTL3. In certain embodiments, administration of a conjugated antisense compound or composition treats, ameliorates, or delays the progression of a cardiovascular and/or metabolic disease.

Certain embodiments disclosed herein provide a method of treating a cardiovascular and/or metabolic disease, comprising: identifying a human having a cardiovascular and/or metabolic disease, and a cardiovascular and/or metabolic disease administering to said human a therapeutically effective amount of any of the conjugated antisense compounds or compositions disclosed herein to treat said human for

Certain embodiments provide conjugated antisense compounds and compositions described herein for use in therapy. In certain embodiments, the therapy is administered to treat, prevent, or delay the progression of a disease associated with ANGPTL3. In certain embodiments, the therapy is administered to treat, prevent, or delay the progression of a disease associated with elevated ANGPTL3.

In certain embodiments, the disease is a cardiovascular and/or metabolic disease, disorder or condition. In certain embodiments, metabolic and/or cardiovascular diseases include, but are not limited to, obesity, diabetes, insulin resistance, atherosclerosis, dyslipidemia, lipodystrophy, coronary heart disease, non-alcoholic fatty liver disease (NAFLD), non-alcoholic fats. hepatitis (NASH) hyperlipidemia or metabolic syndrome, or a combination thereof. The dyslipidemia may be hyperlipidemia. The hyperlipidemia can be combined hyperlipidemia, combined hyperlipidemia (CHL), hypercholesterolemia, hypertriglyceridemia, or both hypercholesterolemia and hypertriglyceridemia. Combined hyperlipidemia may be familial or non-familial. The hypercholesterolemia may be familial allogeneic hypercholesterolemia (HoFH), familial heterozygous hypercholesterolemia (HeFH). ultrahypertriglyceridemia familial chylomicronemia syndrome (FCS) or hyperlipoproteinemia type IV. NAFLD may be hepatic steatosis or steatohepatitis. The diabetes may be type 2 diabetes or type 2 diabetes with dyslipidemia. Insulin resistance can be insulin resistance along with dyslipidemia.

In certain embodiments, a conjugated antisense compound or composition disclosed herein is designated as a first agent, and the method or use disclosed herein further comprises administering a second agent. In certain embodiments, the first agent and the second agent are co-administered. In certain embodiments, the first agent and the second agent are co-administered or administered concurrently.

In certain embodiments, the second agent is a glucose-lowering agent. Glucose lowering agents include, but are not limited to, therapeutic lifestyle changes, PPAR agonists, dipeptidyl peptidase (IV) inhibitors, GLP-1 analogs, insulin or insulin analogs, insulin secretagogues, SGLT2 inhibitors, human amylin analogs, biguanides, an alpha-glucosidase inhibitor, or a combination thereof. Glucose-lowering agents may include, but are not limited to, metformin, sulfonylureas, rosiglitazone, meglitinide, thiazolidinediones, alpha-glucosidase inhibitors, or combinations thereof. The sulfonylurea may be acetohexamide, chlorpropamide, tolbutamide, tolazamide, glimepiride, glipizide, glyburide, or gliclazide. The meglitinide may be nateglinide or repaglinide. The thiazolidinedione may be pioglitazone or rosiglitazone. Alpha-glucosidase may be acarbose or miglitol.

In certain embodiments, the second agent is a lipid-lowering therapy. In certain embodiments, lipid lowering therapies may include, but are not limited to: therapeutic lifestyle changes, HMG-CoA reductase inhibitors, cholesterol absorption inhibitors, MTP inhibitors (eg, small molecules, polypeptides, antibodies or antisense compound (targeted to MTP), ApoB inhibitor (e.g. small molecule, polypeptide, antibody or antisense compound (targeted to ApoB)), ApoC3 inhibitor (e.g. small molecule, polypeptide, antibody or antisense compound (e.g. small molecule, polypeptide, antibody or antisense compound (targeted to ApoC3)) targeted)), PCSK9 inhibitors (eg small molecule, polypeptide, antibody or antisense compound (targeted to PCSK9)), CETP inhibitor (eg small molecule, polypeptide, antibody or antisense compound (targeted to CETP)) , fibrate, beneficial oils (eg krill or fish oil (eg Vascepa ^R ), flaxseed oil, or other omega-3 fatty acid rich oils such as α-linolenic acid (ALA), docosahexae old acid (DHA) or eicosapentaenoic acid (EPA)), or any combination thereof. The HMG-CoA reductase inhibitor may be atorvastatin, rosuvastatin, fluvastatin, lovastatin, pravastatin, or simvastatin. The cholesterol absorption inhibitor may be ezetimibe. The fibrate may be fenofibrate, bezafibrate, ciprofibrate, clofibrate, gemfibrozil, or the like.

In certain embodiments, administration comprises parenteral administration. In certain embodiments, administration comprises subcutaneous administration.

In certain embodiments, administering a conjugated antisense compound disclosed herein results in a decrease in lipid levels, including triglyceride levels, cholesterol levels, insulin resistance, glucose levels, or combinations thereof. One or more of the above levels is independently at least 5%, at least 10%, at least 20%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65% %, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95%. Administration of the conjugated antisense compound may result in enhanced insulin sensitivity or hepatic insulin sensitivity. Administration of the conjugated antisense compounds disclosed herein can result in atherosclerotic plaque, obesity, reduction of glucose, lipids, glucose tolerance, cholesterol, or enhancement of insulin sensitivity or any combination thereof.

Certain embodiments provide for the use of a conjugated antisense compound described herein in the manufacture of a medicament for treating, ameliorating, delaying or preventing one or more of the diseases associated with ANGPTL3. Certain embodiments provide the use of a conjugated antisense compound as described herein in the manufacture of a medicament for treating, ameliorating, or preventing one or more of a metabolic disease or a cardiovascular disease.

Certain embodiments provide a kit for treating, preventing, or ameliorating one or more of a metabolic disease or a cardiovascular disease as described herein, said kit comprising: a) a conjugated antisense as described herein compound; and optionally b) an additional agent or therapy as described herein. The kit may further include instructions or a label for using the kit to treat, prevent, or ameliorate one or more of a metabolic disease or a cardiovascular disease.

antisense compounds

Oligomeric compounds include, but are not limited to, oligonucleotides, oligonucleotides, oligonucleotide analogs, oligonucleotide mimetics, antisense compounds, antisense oligonucleotides, and siRNA. An oligomeric compound may be "antisense" to a target nucleic acid, meaning that it can undergo hybridization to a target nucleic acid via hydrogen bonding.

In certain embodiments, an antisense compound, when written in the 5' to 3' direction, has a nucleobase sequence that comprises the reverse complement of a target segment of a target nucleic acid to which it is targeted. In certain such embodiments, the antisense oligonucleotide, when written in the 5' to 3' direction, has a nucleobase sequence comprising the reverse complement of the target segment of the target nucleic acid to which it is targeted.

In certain embodiments, an antisense compound targeted to an ANGPTL3 nucleic acid is between 10 and 30 nucleotides in length. In other words, antisense compounds are 10 to 30 linked nucleobases. In other embodiments, the antisense compound comprises a modified oligonucleotide consisting of 8 to 80, 10 to 80, 12 to 50, 12 to 30, 15 to 30, 18 to 24, 19 to 22, or 20 linked nucleobases. In certain embodiments, the antisense compound has a length of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27 , 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52 , 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77 , 78, 79, or 80 linked nucleobases or modified oligonucleotides consisting of a range defined by any two of the above values.

In certain embodiments, the antisense compound comprises a shortened or truncated modified oligonucleotide. A shortened or truncated modified oligonucleotide may have a single nucleoside deleted from the 5' end (5' truncation), or alternatively from the 3' end (3' truncated). A shortened or truncated oligonucleotide may have two nucleosides deleted from the 5' end, or alternatively may have two subunits deleted from the 3' end. Alternatively, deleted nucleosides may be dispersed via modified oligonucleotides, for example, in antisense compounds having one or more nucleosides deleted from the 5' end and one or more nucleosides deleted from the 3' end. have.

When a single additional nucleoside is present in an elongated oligonucleotide, the additional nucleoside may be located at the 5' or 3' end of the oligonucleotide. When two or more additional nucleosides are present, the added nucleosides are, for example, 2 added to the 5' end (5' addition), or alternatively, the 3' end (3' addition) of the oligonucleotide may be adjacent to each other in an oligonucleotide having two nucleosides. Alternatively, the added nucleoside may be, for example, one or more nucleosides added to the 5' end and one or more nucleosides added to the 3' end and/or one or more nucleosides added to the central portion. In an oligonucleotide having a seed, it can be dispersed through an antisense compound.

It is possible to increase or decrease the length of an antisense compound, such as an antisense oligonucleotide, and/or introduce mismatched bases without abrogating activity. See, for example, Woolf et al. See, for example, Woolf et al. (Proc. Natl. Acad. Sci. USA 89:7305-7309, 1992), a series of antisense oligonucleotides 13-25 nucleobases in length were tested for their ability to cleave target RNA in an oocyte infusion model. induced Antisense oligonucleotides 25 nucleobases in length with 8 or 11 mismatch bases near the end of the antisense oligonucleotide were capable of direct specific cleavage of the target mRNA, although fewer than antisense oligonucleotides containing no mismatch. . Likewise, target specific cleavage was achieved using 13 nucleobase antisense oligonucleotides, including those with 1 or 3 mismatches.

Gautschi et al. (J. Natl. Cancer Inst. 93:463-471, March 2001) demonstrated 100% complementarity to bcl-2 mRNA to reduce in vitro and in vivo expression of both bcl-2 and bcl-xL. and demonstrated the ability of oligonucleotides with three mismatches to bcl-xL mRNA. Moreover, these oligonucleotides demonstrated potent anti-tumor activity in vivo.

Maher and Dolnick (Nuc. Acid. Res. 16:3341-3358, 1988) described the two or three sequences of each tandem antisense oligonucleotide for its ability to block translation of human DHFR in a rabbit reticulocyte assay. A series of tandem 14 nucleobase antisense oligonucleotides and 28 and 42 nucleobase antisense oligonucleotides consisting of were tested. Each of the three 14 nucleobase antisense oligonucleotides alone was able to inhibit translation at a more moderate level than the 28 or 42 nucleobase antisense oligonucleotides.

Specific antisense compound motifs and mechanisms

In certain embodiments, antisense compounds are arranged in a pattern, or motif, to confer antisense compound properties such as enhanced inhibitory activity, increased binding affinity to a target nucleic acid, or resistance to degradation by nucleases in vivo. It has chemically modified subunits.

Chimeric antisense compounds typically contain at least one region modified to confer increased resistance to nuclease degradation, increased cellular uptake, increased binding affinity of the target nucleic acid, and/or increased inhibitory activity. . The second region of the chimeric antisense compound may confer another desired property and serves as a replacement for, for example, the cellular endonuclease RNase H, which cleaves the RNA strand of the RNA:DNA duplex.

Antisense activity can result from any mechanism involving hybridization of an antisense compound (eg, an oligonucleotide) with a target nucleic acid, which hybridization ultimately results in a biological effect. In certain embodiments, the amount and/or activity of the target nucleic acid is modulated. In certain embodiments, the amount and/or activity of the target nucleic acid is reduced. In certain embodiments, hybridization of an antisense compound to a target nucleic acid does not result in degradation of the target nucleic acid. In certain embodiments, hybridization of an antisense compound to a target nucleic acid does not result in degradation of the target nucleic acid. In certain such embodiments, the presence of an antisense compound hybridized with a target nucleic acid (occupancy) results in modulation of antisense activity. In certain embodiments, antisense compounds having specific chemical motifs or patterns of chemical modifications are particularly suitable for utilizing one or more mechanisms. In certain embodiments, antisense compounds function through more than one mechanism and/or function through an unexplained mechanism. Accordingly, the antisense compounds described herein are not limited by a particular mechanism.

Antisense mechanisms include, but are not limited to: RNase H mediated antisense; RNAi mechanisms that utilize the RISC pathway and include, but are not limited to, siRNA, ssRNA and microRNA mechanisms; and occupancy-based mechanisms. Certain antisense compounds may act through more than one such mechanism and/or through additional mechanisms.

RNaseH - mediated antisense

In certain embodiments, antisense activity results at least in part from degradation of the target RNA by RNase H. RNase H is a cellular endonuclease that cleaves the RNA strand of the RNA:DNA duplex. Single-stranded antisense compounds that are "DNA-like" are known in the art to induce RNase H activity in mammalian cells. Thus, an antisense compound comprising at least a portion of a DNA or DNA-like nucleoside can activate RNase H, which cleaves the target nucleic acid. In certain embodiments, antisense compounds utilizing RNase H comprise one or more modified nucleosides. In certain embodiments, such antisense compounds comprise a block of at least one of 1 to 8 modified nucleosides. In certain such embodiments, the modified nucleoside does not support RNase H activity. In certain embodiments, such antisense compounds are gapmers as described herein. In certain such embodiments, the gap of the gapmer comprises a DNA nucleoside. In certain such embodiments, the gap of the gapmer comprises a DNA-like nucleoside. In certain such embodiments, the gap of the gapmer comprises a DNA nucleoside and a DNA-like nucleoside.

Certain antisense compounds having a gapmer motif are considered to be chimeric antisense compounds. An inner region having a plurality of nucleotides supporting RNaseH cleavage in the gapmer is disposed between an outer region having a plurality of nucleotides chemically distinct from the nucleoside of the inner region. In the case of antisense oligonucleotides having a gapmer motif, the gap segment is generally used as a substrate for endonuclease cleavage, but the wing segment contains modified nucleosides. In certain embodiments, regions of a gapmer are differentiated by the type of sugar moiety comprising each distinct region. The type of sugar moiety used to differentiate a region of a gapmer may in some embodiments include: β-D-ribonucleoside, β-D-deoxyribonucleoside, 2′-modification nucleosides (such 2'-modified nucleosides may include 2'-MOE and 2'-O-CH ₃ , among others), and bicyclic sugar modified nucleosides (such bicyclic Sugar modified nucleosides may include those with limited ethyl). In certain embodiments, the nucleosides in the wing may include several modified sugar moieties, including, for example, 2'-MOE and bicyclic sugar moieties such as restricted ethyl or LNA. In certain embodiments, wings may include several modified and unmodified sugar moieties. In certain embodiments, a wing may comprise various compositions of 2'-MOE nucleosides, bicyclic sugar moieties such as restricted ethyl nucleosides or LNA nucleosides, and 2'-deoxynucleosides.

Each distinct region may contain uniform sugar moieties, variants, or alternating sugar moieties. The wing-gap-wing motif is frequently described as "XYZ", where "X" represents the length of the 5'-wing, "Y" represents the length of the gap, and "Z" represents the length of the 3'-wing. indicates. “X” and “Z” may include moieties per uniform, variant, or alternation. In certain embodiments, "X" and "Y" may include one or more 2'-deoxynucleosides. "Y" may include a 2'-deoxynucleoside. As used herein, a gapmer described as "X-Y-Z" has a configuration such that the cap is disposed immediately adjacent to each of the 5'-wing and 3' wing. Thus, no intervening nucleotides are present between the 5'-wing and the gap, or between the gap and the 3'-wing. Any of the antisense compounds described herein may have a gapmer motif. In certain embodiments, “X” and “Z” are the same, and in other embodiments, y is different. In certain embodiments, “Y” is 8 to 15 nucleosides. X, Y, or Z is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30 or any more nucleosides.

In certain embodiments, the antisense compound is targeted to an ANGPTL3 nucleic acid and has a gapmer motif in which the gap consists of 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 linked nucleosides.

In certain embodiments, antisense oligonucleotides have a sugar motif described in Formula A as follows: (J) _m -(B) _n -(J) _p -(B) _r -(A) _t -(D) _g -(A) _v -(B) _w -(J) _x -(B) _y -(J) _z

In the above formula,

each A is independently a 2'-substituted nucleoside;

each B is independently a bicyclic nucleoside;

each J is independently a 2'-substituted nucleoside or a 2'-deoxynucleoside;

each D is a 2'-deoxynucleoside;

m is 0 to 4; n is 0 to 2; p is 0 to 2; r is 0 to 2; t is 0 to 2; v is 0 to 2; w is 0 to 4; x is 0 to 2; y is 0 to 2; z is 0 to 4; G is 6-14;

only,

at least one of m, n, and r is other than 0;

at least one of w and y is other than 0;

the sum of m, n, p, r, and t is 2 to 5; and

The sum of v, w, x, y, and z is 2 to 5.

RNAi compounds

In certain embodiments, the antisense compound is an interfering RNA compound (RNAi), including double-stranded RNA compounds (also called single-interfering RNA or siRNA) and single-stranded RNAi compounds (or ssRNA). Such compounds act at least in part via the RISC pathway to degrade and/or sequester target nucleic acids (thus include microRNA/microRNA-mimic compounds). In certain embodiments, antisense compounds include modifications to make them particularly suitable for such a mechanism.

One. ssRNA compound

In certain embodiments, antisense compounds, including those particularly suitable for use as single-stranded RNAi compounds (ssRNA), comprise a modified 5'-end. In certain such embodiments, the 5'-end comprises a modified phosphate moiety. In certain embodiments, such modified phosphates are stabilized (eg, resistant to degradation/cleavage compared to unmodified 5'-phosphate). In certain embodiments, such a 5'-terminal nucleoside stabilizes the 5'-in moiety. Certain modified 5'-terminal nucleosides can be found in the art, for example WO/2011/139702.

In certain embodiments, the 5'-nucleoside of the ssRNA compound has the formula IIc:

In the above formula,

T ₁ is optionally a protected phosphorus moiety;

T ₂ is an internucleoside linking group linking the compound of IIc to the oligomeric compound;

A has one of the formulas:

Q ₁ and Q ₂ are each, independently, H, halogen, C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted C ₁ -C ₆ alkoxy, C ₂ - C ₆ alkenyl, substituted C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, substituted C ₂ -C ₆ alkynyl or N(R ₃ )(R ₄ );

Q ₃ is O, S, N(R ₅ ) or C(R ₆ )(R ₇ );

each of R ₃ , R ₄ R ₅ , R ₆ and R ₇ is independently H, C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy;

M ₃ is O, S, NR ₁₄ , C(R ₁₅ )(R ₁₆ ), C(R ₁₅ )(R ₁₆ )C(R ₁₇ )(R ₁₈ ), C(R ₁₅ )=C(R ₁₇ ) , OC(R ₁₅ )(R ₁₆ ) or OC(R ₁₅ )(Bx ₂ );

R ₁₄ is H, C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, substituted C ₂ - C ₆ alkenyl, C ₂ -C ₆ alkynyl or substituted C ₂ -C ₆ alkynyl;

R ₁₅ , R ₁₆ , R ₁₇ and R ₁₈ are each, independently, H, halogen, C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, substituted C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or substituted C ₂ -C ₆ alkynyl;

Bx ₁ is a heterocyclic base moiety.

or if Bx ₂ is present, Bx ₂ is a heterocyclic base moiety and Bx ₁ is H, halogen, C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, substituted C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or substituted C ₂ -C ₆ alkynyl;

J ₄ , J ₅ , J ₆ and J ₇ are each, independently, H, halogen, C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, substituted C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or substituted C ₂ -C ₆ alkynyl;

or J ₄ forms a bridge with one of J ₅ or J ₇ wherein said bridge comprises 1-3 linked divalent radical groups selected from: O, S, NR ₁₉ , C(R ₂₀ )(R ₂₁ ), C(R ₂₀ )=C(R ₂₁ ), C[=C(R ₂₀ )(R ₂₁ )], and the other two of C(=O), J ₅ , J ₆ and J _{7 , respectively,} independently H, halogen, C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, substituted C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or substituted C ₂ -C ₆ alkynyl;

each of R ₁₉ , R ₂₀ and R ₂₁ is, independently, H, C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted C ₁ -C ₆ alkoxy, C ₂ -C ₆ alkenyl, substituted C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or substituted C ₂ -C ₆ alkynyl;

G is H, OH, halogen, or O-[C(R ₈ )(R ₉ )] _n -[(C=O) _m -X ₁ ] _j -Z;

each of R ₈ and R ₉ is, independently, H, halogen, C ₁ -C ₆ alkyl or substituted C ₁ -C ₆ alkyl;

X ₁ is O, S or N(E ₁ );

Z is H, halogen, C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, substituted C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, substituted C ₂ -C ₆ alkynyl or N(E ₂ )(E ₃ );

E ₁ , E ₂ and E ₃ are each, independently, H, C ₁ -C ₆ alkyl or substituted C ₁ -C ₆ alkyl;

n is 1 to about 6;

m is 0 or 1;

j is 0 or 1;

Each substituted group includes one or more optionally protected substituent groups independently selected from: halogen, OJ ₁ , N(J ₁ )(J ₂ ), =NJ ₁ , SJ ₁ , N ₃ , CN, OC(=X ₂ )J ₁ , OC(=X ₂ )-N(J ₁ )(J ₂ ) and C(=X ₂ )N(J ₁ )(J ₂ );

X ₂ is O, S or NJ ₃ ;

each of J ₁ , J ₂ and J ₃ is, independently, H or C ₁ -C ₆ alkyl; and

if j is 1, then Z is halogen or other than _{N(E 2} )(E _{3 );} and

The oligomeric compound comprises from 8 to 40 monomeric subunits and is hybridizable to at least a portion of the target nucleic acid.

In certain embodiments, M ₃ is O, CH=CH, OCH ₂ or OC(H)(Bx ₂ ). In certain embodiments, M ₃ is O.

each of J ₄ , J ₅ , J ₆ and J ₇ is, independently, H. In certain embodiments, J ₄ forms a bridge with either J _{5 or J 7 .}

In certain embodiments, A has one of the formulas:

In the above formula,

Q ₁ and Q ₂ are each, independently, H, halogen, C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy or substituted C ₁ -C ₆ alkoxy. In certain embodiments, Q ₁ and Q ₂ are each H. In certain embodiments, Q ₁ and Q ₂ are each, independently, H or halogen. In certain embodiments, Q ₁ and Q ₂ are H and the _{other of Q 1} and Q ₂ is F, CH ₃ or OCH ₃ .

In certain embodiments, T ₁ has the formula:

In the above formula,

each of R _a and R _c is, independently, protected hydroxyl, protected thiol, C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted C ₁ -C ₆ alkoxy, protected amino or substituted amino; and

R _b is O or S. In certain embodiments, R _b is O and _{each of R a} and R _c is, independently, OCH ₃ , OCH ₂ CH _{3 ,} or CH(CH ₃ ) ₂ .

In certain embodiments, G is halogen, OCH ₃ , OCH ₂ F, OCHF ₂ , OCF ₃ , OCH ₂ CH ₃ , O(CH ₂ ) ₂ F, OCH ₂ CHF ₂ , OCH ₂ CF ₃ , OCH ₂ -CH= CH ₂ , O(CH ₂ ) ₂ -OCH ₃ , O(CH ₂ ) ₂ -SCH ₃ , O(CH ₂ ) ₂ -OCF ₃ , O(CH ₂ ) ₃ -N(R ₁₀ )(R ₁₁ ), O(CH ₂ ) ₂ -ON(R ₁₀ )(R ₁₁ ), O(CH ₂ ) ₂ -O(CH ₂ ) ₂ -N(R ₁₀ )(R ₁₁ ), OCH ₂ C(=O)-N (R ₁₀ )(R ₁₁ ), OCH ₂ C(=O)-N(R ₁₂ )-(CH ₂ ) ₂ -N(R ₁₀ )(R ₁₁ ) or O(CH ₂ ) ₂ -N(R _{12 )} )-C(=NR ₁₃ )[N(R ₁₀ )(R ₁₁ )], wherein R ₁₀ , R ₁₁ , R ₁₂ and R ₁₃ are each, independently, H or C ₁ -C ₆ alkyl. In certain embodiments, G is halogen, OCH ₃ , OCF ₃ , OCH ₂ CH ₃ , OCH ₂ CF ₃ , OCH ₂ -CH=CH ₂ , O(CH ₂ ) ₂ -OCH ₃ , O(CH ₂ ) ₂ - O(CH ₂ ) ₂ -N(CH ₃ ) ₂ , OCH ₂ C(=O)-N(H)CH ₃ , OCH ₂ C(=O)-N(H)-(CH ₂ ) ₂ -N( CH ₃ ) ₂ or OCH ₂ -N(H)-C(=NH)NH ₂ . In certain embodiments, G is F, OCH ₃ or O(CH ₂ ) ₂ —OCH ₃ . In certain embodiments, G is O(CH ₂ ) ₂ -OCH ₃ .

In certain embodiments, the 5'-terminal nucleoside has the formula IIe:

In certain embodiments, antisense compounds, including those particularly suitable for ssRNA, comprise one or more types of modified sugar moieties and/or naturally occurring sugar moieties or regions thereof prepared with oligonucleotides in a defined pattern or sugar modification motif. includes Such motifs may include any of the sugar modifications discussed herein and/or other known sugar modifications.

In certain embodiments, such motifs may include any of the sugar modifications discussed herein and/or other known sugar modifications. In certain such embodiments, each nucleoside of said region comprises the same RNA-like sugar modification. In certain embodiments, each nucleoside of said region is a 2'-F nucleoside. In certain embodiments, each nucleoside of said region is a 2'-OMe nucleoside. In certain embodiments, each nucleoside of said region is a 2'-MOE nucleoside. In certain embodiments, each nucleoside of said region is a cEt nucleoside. In certain embodiments, each nucleoside of said region is an LNA nucleoside. In certain embodiments, the homogeneous region constitutes all or essentially all oligonucleotides. In certain embodiments, a region constitutes the entire oligonucleotide except for one to four terminal nucleosides.

In certain embodiments, the oligonucleotide comprises one or more regions of alternating sugar modifications, wherein the nucleoside alternates between two nucleotides having a first type of sugar modification and a nucleotide having a second type of sugar modification. In certain embodiments, both types of nucleosides are RNA-like nucleosides. In certain embodiments, the alternating nucleosides are selected from: 2'-OMe, 2'-F, 2'-MOE, LNA, and cEt. In certain embodiments, the alternating modifications are 2'-F and 2'-OMe. Such regions may be contiguous or may be interrupted by differently modified nucleosides or conjugated nucleosides.

In certain embodiments, the alternating region of each alternating modification consists of a single nucleoside (ie, the pattern is (AB) _x A _y , wherein A is a nucleoside having a sugar modification of a first type and B is a second nucleosides having two types of sugar modifications; X is 1 to 20 and y is 0 or 1. In certain embodiments, one or more alternating regions of the alternating motif comprises more than a single type of nucleoside. For example, an oligonucleotide may comprise a region of any one or more of the following nucleoside motifs:

AABBAA;

ABBABB;

AABAAB;

ABBABAABB;

ABBAAA;

AABABAB;

ABBAAA;

ABBAABBABABAA;

BABBAABBABABAA; or

ABABBAABBABABAA;

wherein A is a nucleoside of a first type and B is a nucleoside of a second type. In certain embodiments, each of A and B is selected from 2'-F, 2'-OMe, BNA, and MOE.

In certain embodiments, oligonucleotides with such alternating motifs also include modified 5' terminal nucleosides, such as those of formula IIc or IIe.

In certain embodiments, the oligonucleotide comprises a region having a 2-2-3 motif. Such areas include the following motifs:

-(A) ₂ -(B) _x -(A) ₂ -(C) _y -(A) ₃ -

wherein A is a modified nucleoside of the first type;

B and C are nucleosides modified differently than A, however, B and C may have the same or different modifications from each other;

X and y are 1 to 15.

In certain embodiments, A is a 2'-OMe modified nucleoside. In certain embodiments, both B and C are 2'-F modified nucleosides. In certain embodiments, A is a 2'-OMe modified nucleoside and both B and C are 2'-F modified nucleosides.

In certain embodiments, the oligonucleotide has the following sugar motifs:

5'-(Q)-(AB) _x A _y -(D) _z

In the above formula,

Q is a nucleoside comprising a stable phosphate moiety. In certain embodiments, Q is a nucleoside having formula IIc or IIe;

A is a modified nucleoside of the first type;

B is a modified nucleoside of a second type;

D is a modified nucleoside comprising a different modification from the nucleoside adjacent to it. Thus, if y is 0, then D must deform differently than B, and if y is 1, then D must deform differently than A. In certain embodiments, D is different from both A and B.

X is 5 to 15;

Y is 0 or 1;

Z is 0 to 4.

In certain embodiments, the oligonucleotide has the following sugar motifs:

5'-(Q)-(A) _x -(D) _z

In the above formula,

Q is a nucleoside comprising a stable phosphate moiety. In certain embodiments, Q is a nucleoside having formula IIc or IIe;

A is a modified nucleoside of the first type;

D is a modified nucleoside comprising a different modification from A.

X is 11 to 30;

Z is 0 to 4.

In certain embodiments, A, B, C, and D of the motifs are selected from: 2'-OMe, 2'-F, 2'-MOE, LNA, and cEt. In certain embodiments, D represents a terminal nucleoside. In certain embodiments, such terminal nucleosides are not designed to hybridize to the target nucleic acid (but one or more do hybridize by chance). In certain embodiments, the nucleobase of each D nucleoside is adenine, regardless of the identity of the nucleobase at the corresponding position in the target nucleic acid. In certain embodiments the nucleobase of each D nucleoside is thymine.

In certain embodiments, antisense compounds, including those particularly suitable for use as ssRNA, comprise modified internucleoside linkages arranged along an oligonucleotide or region thereof in a defined pattern or modified internucleoside linkage motif. In certain embodiments, the oligonucleotide comprises regions with alternating internucleoside linkage motifs. In certain embodiments, the oligonucleotide comprises regions of uniformly modified internucleoside linkages. In certain such embodiments, the oligonucleotide comprises regions uniformly linked by phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotides are homogeneously linked by phosphorothioate internucleoside linkages. In certain embodiments, each internucleoside linkage of the oligonucleotide is selected from a phosphodiester and a phosphorothioate. In certain embodiments, each internucleoside linkage of the oligonucleotide is selected from phosphodiester and phosphorothioate and at least one internucleoside linkage is phosphorothioate.

In certain embodiments, the oligonucleotide comprises at least 6 phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least 8 phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least 10 phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least 6 consecutive phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least 8 consecutive phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least 10 consecutive phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least one 12 consecutive phosphorothioate internucleoside linkages. In certain such embodiments, at least one such block is located at the 3' end of the oligonucleotide. In certain such embodiments, at least one such block is located within three nucleosides of the 3' end of the oligonucleotide.

Oligonucleotides having any of the various sugar motifs described herein can have any linking motif. For example, oligonucleotides, including but not limited to those described above, may have a linking motif selected from the non-limiting table below:

2. siRNA compound

In certain embodiments, the antisense compound is a double-stranded RNAi compound (siRNA). In this embodiment, one or both strands may contain any of the modifying motifs described above for ssRNA. In certain embodiments, the ssRNA compound may be unmodified RNA. In certain embodiments, siRNA compounds may comprise unmodified RNA nucleosides and modified internucleoside linkages.

Several embodiments relate to double-stranded compositions, wherein each strand comprises a motif defined by the position of one or more modified or unmodified nucleosides. In certain embodiments, compositions are provided comprising first and second oligomeric compounds that fully or at least partially hybridize to form a duplex region and further comprising a region that is complementary to and hybridizes to a nucleic acid target. The composition comprises a first oligomeric compound that is an antisense strand having complete or partial complementarity to a nucleic acid target and a second oligomeric compound that is a sense strand having one or more regions of complementarity to the first oligomeric compound to form at least one duplex region it is suitable to do

Compositions of several embodiments modulate gene expression by hybridizing to a nucleic acid target and thereby losing its normal function. In certain embodiments, the target nucleic acid is ANGPTL3. In certain embodiments, the degradation of targeted ANGPTL3 is facilitated by an activated RISC complex formed with a composition of the invention.

Several embodiments relate to double-stranded compositions, wherein one of the strands is useful, for example, to effect preferential loading of the opposite strand into the RISC (or cleavage) complex. The composition is useful for targeting a selected nucleic acid molecule and modulating the expression of one or more genes. In some embodiments, the composition of the present invention hybridizes to a portion of the target RNA, resulting in loss of normal function of the target RNA.

Certain embodiments relate to double-stranded compositions, wherein both strands comprise a hemimer motif, a fully modified motif, a positionally modified motif, or an alternating motif. Each strand of the composition of the invention can be modified to fulfill a specific role, for example, in the siRNA pathway. The use of the same motif with different motifs on each strand or with different chemical modifications on each strand makes it possible to target the antisense strand to the RISC complex while inhibiting incorporation of the sense strand. Within this model, each strand is independently modified so that it can be enhanced for its specific role. The antisense strand can be modified at the 5' end to enhance its role in one region of RISC and the 3' end can be modified differently to enhance its role in different regions of RISC.

The double-stranded oligonucleotide molecule may be a double-stranded polynucleotide molecule comprising self-complementary sense and antisense regions, wherein the antisense region comprises a nucleotide sequence complementary to a nucleotide sequence in its target nucleic acid molecule or portion thereof, and wherein the sense The region has a nucleotide sequence corresponding to the target nucleic acid sequence or portion thereof. Double-stranded oligonucleotide molecules can be assembled from two separate oligonucleotides, one strand being the sense strand and the other being the antisense strand, wherein the antisense and sense strands are self-complementary (i.e., each strand is nucleotide sequence complementary to the nucleotide sequence in the strand); For example, the antisense strand and the sense strand may have a double-stranded or double-stranded structure, e.g., the double-stranded region has from about 15 to about 30, e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 base pairs; The antisense strand comprises a nucleotide sequence complementary to a nucleotide sequence within a target nucleic acid molecule or a portion thereof, and the sense strand comprises a target nucleic acid sequence or portion thereof (e.g., about 15 to about 25 of a double-stranded oligonucleotide molecule). more than one nucleotide is complementary to the target nucleic acid or portion thereof). Alternatively, double-stranded oligonucleotides are assembled from a single oligonucleotide, wherein the self-complementary sense and antisense regions of the siRNA are linked by nucleic acid-based or non-nucleic acid-based linker(s).

The double-stranded oligonucleotide may be a polynucleotide having a duplex, asymmetric duplex, hairpin or asymmetric hairpin secondary structure, having self-complementary sense and antisense regions, wherein the antisense region is a separate target nucleic acid molecule or and a nucleotide sequence complementary to a nucleotide sequence in a portion thereof, wherein the sense region has a nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof. The double-stranded oligonucleotide may be a circular single-stranded polynucleotide having two or more loop structures and a stem comprising self-complementary sense and antisense regions, wherein the antisense region is a nucleotide sequence in a target nucleic acid molecule or portion thereof. and wherein the sense region has a nucleotide sequence corresponding to a target nucleic acid sequence or a portion thereof, wherein the circular polynucleotide is processed in vivo or in vitro to generate an active siRNA molecule capable of mediating RNAi. can be

In certain embodiments, the double-stranded oligonucleotide comprises separate sense and antisense sequences or regions, wherein the sense and antisense regions are covalently linked by nucleotide or non-nucleotide linker molecules or alternatively, as is known in the art. They are non-covalently linked by ionic interactions, hydrogen bonding, van der Waals interactions, hydrophobic interactions and/or stacking interactions. In certain embodiments, the double-stranded oligonucleotide comprises a nucleotide sequence complementary to the nucleotide sequence of the target gene. In another embodiment, the double-stranded oligonucleotide interacts with a nucleotide sequence of a target gene in a manner that inhibits expression of the target gene.

As used herein, double-stranded oligonucleotides need not be limited to those molecules containing only RNA, but further include chemically modified nucleotides and non-nucleotides. In certain embodiments, the short interfering nucleic acid molecule is free of 2'-hydroxy (2'-OH) containing nucleotides. In certain embodiments, short interfering nucleic acids optionally do not include any ribonucleotides (eg, nucleotides with a 2′-OH group). However, such double-stranded oligonucleotides that do not require the presence of intramolecular ribonucleotides to support RNAi may contain one or more nucleotides with an attached linker or linkers or other attached or associated group, moiety or 2'-OH group. It may have a chain containing Optionally, the double-stranded oligonucleotide may comprise ribonucleotides at about 5, 10, 20, 30, 40, or 50% of the nucleotide positions. The term siRNA as used herein refers to sequence specific RNAi, e.g., short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), short hairpin RNA (shRNA), short interfering oligonucleotides. , short interfering nucleic acids, short interfering modified oligonucleotides, chemically modified siRNAs, post-transcriptional gene silencing RNA (ptgsRNA), and other terms used to describe nucleic acid molecules capable of mediating . Additionally, the term RNAi as used herein is meant to be equivalent to other terms used to describe sequence specific RNA interference such as post-transcriptional gene silencing, translational repression, or epigenetics. For example, double-stranded oligonucleotides can be used to epigenetically silence genes at both the post-transcriptional level and the pre-transcriptional level. In a non-limiting example, epigenetic regulation of gene expression by an siRNA molecule of the invention may result from siRNA-mediated modifications of chromatin structure or methylation pattern to alter gene expression (see, e.g. , Verdel et al., (2004) Science 303:672-676; Pal-Bhadra et al. 2004, Science, 303, 669-672; Allshire, 2002, Science, 297, 1818-1819; Volpe et al., 2002 , Science, 297, 1833-1837; Jenuwein, 2002, Science, 297, 2215-2218; and Hall et al., 2002, Science, 297, 2232-2237).

The compounds and compositions of the various embodiments provided herein are dsRNA-mediated gene silencing or RNAi mechanisms, including, for example, "hairpins" or stem-loop double-stranded RNA effector molecules, e.g., "hairpins" or stem- It is contemplated that ANGPTL3 can be targeted by a stem-loop double-stranded RNA effector molecule, wherein a double-stranded conformation with a single RNA strand having a self-complementary sequence or a double comprising two separate strands of RNA A helical dsRNA effector molecule can be inferred. In various embodiments, the dsRNA consists entirely of ribonucleotides, e.g., in WO 00/63364, filed April 19, 2000, or U.S. Serial No. 60/130,377, filed April 21, 1999 It consists of mixtures of ribonucleotides and deoxynucleotides, such as RNA/DNA hybrids, as described by The dsRNA or dsRNA effector molecule may be a single molecule having a region of self-complementarity such that nucleotides in one segment of the molecule base pair with nucleotides in another segment of the molecule. In various embodiments, a dsRNA of a single molecule consists entirely of ribonucleotides or comprises a ribonucleotide region complementary to a deoxyribonucleotide region. Alternatively, the dsRNA may comprise two different strands having regions of complementarity to each other.

In various embodiments, both strands consist entirely of ribonucleotides, one strand consists entirely of ribonucleotides and one strand consists entirely of deoxyribonucleotides, or one or both strands consist entirely of ribonucleotides and deoxyribonucleotides. Contains a mixture of oxyribonucleotides. In certain embodiments, the regions of complementarity are at least 70, 80, 90, 95, 98, or 100% complementary to each other and to the target nucleic acid sequence. In certain embodiments, the dsRNA region that exists in the double-stranded conformation is at least 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 50, 75,100, 200, 500, 1000, 2000 or 5000 nucleotides or all nucleotides as cDNA or other target nucleic acid sequences are provided as dsRNAs. In some embodiments, the dsRNA does not contain any single-stranded regions, such as single-stranded ends, or the dsRNA is a hairpin. In other embodiments, the dsRNA has one or more single-stranded regions or overhangs. In certain embodiments, the RNA/DNA hybrid comprises a DNA strand or region that is an antisense strand or region (e.g., has at least 70, 80, 90, 95, 98, or 100% complementarity with a target nucleic acid) and a sense strand or RNA strand or region that is a region (eg, has at least 70, 80, 90, 95, 98, or 100% identity to a target nucleic acid), and vice versa.

In various embodiments, RNA/DNA hybrids are synthesized from the methods described herein or those described in WO 00/63364, filed April 19, 2000, or U.S. Serial No. 60/130,377, filed April 21, 1999; It is prepared in vitro using the same enzymatic or chemical synthetic methods. In another embodiment, the DNA strand synthesized in vitro is complexed with an RNA strand prepared in vivo or in vitro prior to, after or concurrent with transformation of the DNA strand into a cell. In another embodiment, the dsRNA is a single circular nucleic acid containing sense and antisense regions, or the dsRNA comprises a circular nucleic acid and a second circular nucleic acid or a linear nucleic acid (see, e.g., on Apr. 19, 2000). filed WO 00/63364 or U.S. Serial No. 60/130,377, filed April 21, 1999). Exemplary circular nucleic acids include a lariat structure, wherein the free 5' phosphoryl group of a nucleotide is linked to the 2' hydroxyl group of another nucleotide in a loop back fashion.

In other embodiments, the dsRNA comprises one or more modified nucleotides, wherein the 2' position in the sugar contains a halogen (eg, a fluorine group) or the corresponding 2' position contains a hydrogen or hydroxyl group contains an alkoxy group (eg, a methoxy group) that increases the half-life of the dsRNA in vitro or in vivo compared to its corresponding dsRNA. In another embodiment, the dsRNA comprises one or more linkages between adjacent nucleotides other than naturally occurring phosphodiester linkages. Examples of such linkages include phosphoramide, phosphorothioate and phosphorodithioate linkages. A dsRNA may also be a chemically modified nucleic acid molecule taught in US Pat. No. 6,673,661. In other embodiments, the dsRNA is, e.g., as described by WO 00/63364 or 19, 2000, filed April 19, 2000, or 60/130,377, filed April 21, 1999, U.S. Serial No. Contains one or two capped strands.

In other embodiments, the dsRNA may be any of the at least partial dsRNA molecules disclosed in WO 00/63364, as well as in US Provisional Applications 60/399,998; and dsRNA molecules disclosed in U.S. Provisional Application Nos. 60/419,532, and PCT/US2003/033466, the teachings of which are incorporated herein by reference. Any dsRNA can be expressed in vitro or in vivo using the methods described herein or standard methods such as those described in WO 00/63364.

occupancy

In certain embodiments, antisense compounds are not expected to induce cleavage or target nucleic acid via RNase H or induce cleavage or sequestration via the RISC pathway. In certain such embodiments, antisense activity may result from occupancy, wherein the presence of the hybridized antisense compound disrupts the activity of the target nucleic acid. In certain such embodiments, the antisense compound may comprise uniformly modified or mixed modified and/or modified and unmodified nucleosides.

Target nucleic acid, target region and nucleotide sequence

Nucleotide sequences encoding ANGPTL3 include, but are not limited to, the human sequence set forth in Genbank Accession No. NM_014495.2 (incorporated herein as SEQ ID NO: 1) or Genbank Accession Nos. NT_032977.9 Nucleotides 33032001 to 33046000 (herein in SEQ ID NO: 2). It is understood that the sequence set forth by each SEQ ID NO in the Examples contained herein is independent of any modifications to sugar moieties, internucleoside linkages, or nucleobases. As such, the antisense compound defined by SEQ ID NO: may independently comprise one or more modifications to a sugar moiety, an internucleoside linkage, or a nucleobase. Antisense compounds described by ISIS number (Isis No) represent a combination of nucleobase sequences and motifs.

In certain embodiments, the target region is a structurally defined region of a target nucleic acid. For example, a target region may comprise a 3' UTR, 5' UTR, exon, intron, exon/intron conjugate, coding region, translation initiation region, translation termination region, or other defined nucleic acid region. The structurally defined region for ANGPTL3 can be obtained by accession number from a sequence database such as NCBI, and such information is incorporated herein by reference. In certain embodiments, the target region may encompass a sequence from the 5' target site of one target segment within the target region to the 3' target site of another target segment within the target region.

In certain embodiments, a “target segment” is a smaller, sub-portion of a target region in a nucleic acid. For example, a “target segment” may be a sequence of nucleotides of a target nucleic acid to which one or more antisense compounds are targeted. "5' target site" or "5' stop site" refers to the 5'-most nucleotide of the target segment. "3' target site" or "3' stop site" refers to the 3'-most nucleotide of the target segment.

Targeting involves determining at least one target segment to which the antisense compound hybridizes, thereby resulting in a desired effect. In certain embodiments, the desired effect is a decrease in mRNA target nucleic acid levels. In certain embodiments, the desired effect is a reduction in the level of the protein encoded by the target nucleic acid or a phenotypic change associated with the target nucleic acid.

The target region may contain one or more target segments. Multiple target segments within a target region may be overlapping. Alternatively, they may be non-overlapping. In certain embodiments, target segments within the target region are separated by no more than about 300 nucleotides. In certain embodiments, the target segment within the target region is about 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 or fewer nucleotides on the target nucleic acid, and separated by a number of nucleotides in the range defined by any two of the values. In certain embodiments, target segments within the target region are separated by no more than about 5 nucleotides on the target nucleic acid. In certain embodiments, the target segments are contiguous. Target regions defined by ranges having an initiating nucleic acid that are any of the 5' target sites or 3' target sites listed herein are contemplated.

Suitable target segments can be found within the 5' UTR, coding region, 3' UTR, intron, exon, or exon/intron conjugate. A target segment containing a start codon or a stop codon is also a suitable target segment. Suitable target segments may explicitly exclude certain structurally defined regions such as start codons or stop codons.

Determination of a suitable target segment may include comparison of the sequence of the target nucleic acid to other sequences throughout the genome. For example, the BLAST algorithm can be used to identify regions of similarity among different nucleic acids. Such comparison may prevent selection of sequences of antisense compounds that are capable of hybridizing in a non-specific manner with sequences other than the selected target nucleic acid (ie, non-target or off-target sequences).

There may be a change in the activity of the antisense compound within the active target region (eg, as defined by a percent decrease in the level of the target nucleic acid). In certain embodiments, a decrease in ANGPTL3 mRNA level is indicative of inhibition of ANGPTL3 protein expression. Decreased levels of ANGPTL3 protein also indicate inhibition of target mRNA expression. Moreover, phenotypic changes, such as reduced cholesterol, LDL, triglyceride, or glucose levels, may indicate inhibition of ANGPTL3 mRNA and/or protein expression.

hybridization

In some embodiments, hybridization occurs between an antisense compound disclosed herein and an ANGPTL3 nucleic acid. The most common mechanism of hybridization involves hydrogen bonding between complementary nucleobases of a nucleic acid molecule (e.g., Watson-Crick, Hoogsteen, or inverted Hoogsteen hydrogen bonding). .

Hybridization can occur under variable conditions. Stringent conditions are sequence-dependent and are determined by the nature and composition of the nucleic acid molecule to be hybridized.

Methods for determining whether a sequence can specifically hybridize with a target nucleic acid are known in the art (Sambrooke and Russell, Molecular Cloning: A Laboratory Manual, 3 ^rd Ed., 2001). In certain embodiments, the antisense compounds provided herein are capable of specifically hybridizing with an ANGPTL3 nucleic acid.

complementarity

Antisense compounds and target nucleic acids are formed when a sufficient number of nucleobases of the antisense compound can hydrogen bond with the corresponding nucleobases of the target nucleic acid, thereby resulting in a desired effect (e.g., antisense inhibition of a target nucleic acid, such as an ANGPTL3 nucleic acid). , complementary to each other.

Moreover, the antisense compound can hybridize on one or more segments of the ANGPTL3 nucleic acid such that the intervening or adjacent segments are not involved in a hybridization event (eg, loop structures, mismatches or hairpin structures).

In certain embodiments, an antisense compound provided herein, or a specific portion thereof, is 70%, 75%, 80%, 85%, 86%, 87%, 88 in an ANGPTL3 nucleic acid, target region, target segment, or specific portion thereof. %, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or at least complementary as above. In certain embodiments, an antisense compound provided herein, or a specific portion thereof, comprises one or more sequences of SEQ ID NOs: 1-2 and 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89 %, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or at least complementary as above. The percent complementarity of an antisense compound with a target nucleic acid can be determined using routine methods.

For example, an antisense compound, in which 18 of the nucleobases of the 20 antisense compounds are complementary to the target region and thus specifically hybridize, will exhibit 90 percent complementarity. In this example, residual non-complementary nucleobases may cluster or intersperse with complementary nucleobases, and need not be adjacent to each other or to complementary nucleobases. As such, an antisense compound of 18 nucleobases in length with 4 non-complementary nucleobases flanked by two regions with perfect complementarity to the target nucleic acid will have 77.8% total complementarity to the target nucleic acid, thus will be within the scope of the present invention. The percent complementarity of an antisense compound with a region of a target nucleic acid can be routinely determined using the BLAST program (basic local alignment study tool) and the PowerBLAST program known in the art (Altschul et al., J. Mol. Biol., 1990, 215, 403 410; Zhang and Madden, Genome Res., 1997, 7, 649 656). Percent homology, sequence identity or complementarity can be determined using default settings using the algorithm of Smith and Waterman (Adv. Appl. Math., 1981, 2, 482 489), for example, the Gap program (Wisconsin Sequence). Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, Madison Wis.).

In certain embodiments, an antisense compound provided herein, or a specified portion thereof, is fully complementary (ie, 100% complementary) to a target nucleic acid, or specified portion thereof. For example, an antisense compound may be completely complementary to an ANGPTL3 nucleic acid, or a target region thereof, or a target segment or target sequence. As used herein, "fully complementary" means that each nucleobase of an antisense compound is capable of correct base pairing with the corresponding nucleobase of a target nucleic acid. For example, a 20 nucleobase antisense compound is perfectly complementary to a target sequence that is 400 nucleobases in length as long as there is a corresponding 20 nucleobase portion of the target nucleic acid that is perfectly complementary to the antisense compound. Perfect complementarity can also be used with reference to a specified portion of a first and/or second nucleic acid. For example, a 20 nucleobase portion of a 30 nucleobase antisense compound may be “fully complementary” to a target sequence that is 400 nucleobases in length. The 20 nucleobase portion of the 30 nucleobase oligonucleotide is completely complementary to the target sequence if the target sequence has a corresponding 20 nucleobase portion, wherein each nucleobase is complementary to the 20 nucleobase portion of the antisense compound. to be. At the same time, the total 30 nucleobase antisense compound can also be completely complementary to the target sequence depending on whether the remaining 10 nucleobases of the antisense compound are also complementary to the target sequence.

The position of the non-complementary nucleobase may be at the 5' end or the 3' end of the antisense compound. Alternatively, a non-complementary nucleobase or nucleobases may be at an internal position of the antisense compound. When two or more non-complementary nucleobases are present, they may be contiguous (ie linked) or non-contiguous. In one embodiment, the non-complementary nucleobase is located in the wing segment of the gapmer antisense oligonucleotide.

In certain embodiments, an antisense compound of 10, 12, 13, 14, 15, 16, 17, 18, 19, or 20 in length, or at most that length, comprises a target nucleic acid, such as an ANGPTL3 nucleic acid, or a specified portion thereof. 6 or less, 5 or less, 4 or less, 3 or less, 2 or less, or 1 or less non-complementary nucleobase(s) for

In certain embodiments, the length is 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30; or at most such length, the antisense compound may contain no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 non-complementary nucleobase(s) to a target nucleic acid, such as an ANGPTL3 nucleic acid, or a specified portion thereof. includes

Antisense compounds provided herein also include those that are complementary to a portion of a target nucleic acid. As used herein, “portion” refers to a defined number of contiguous (ie, linked) nucleobases within a region or segment of a target nucleic acid. A “portion” may also refer to a defined number of contiguous nucleobases of an antisense compound. In certain embodiments, the antisense compound is complementary to at least an 8 nucleobase portion of the target segment. In certain embodiments, the antisense compound is complementary to at least a 10 nucleobase portion of the target segment. In certain embodiments, the antisense compound is complementary to at least a 15 nucleobase portion of the target segment. nucleobases in the range defined by at least 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more of the target segment, or any two of these values Antisense compounds complementary to the moiety are also contemplated.

sameness

The antisense compounds provided herein may also have a defined percent identity to a particular nucleotide sequence, SEQ ID NO, or a compound represented by a particular SEQ ID NO: or a portion thereof. As used herein, an antisense compound is identical to a sequence disclosed herein if it has the same nucleobase pairing ability. For example, RNA containing uracil in place of thymidine in the disclosed DNA sequence is considered identical to the DNA sequence, as both uracil and thymidine pair with adenine. Abbreviated and extended versions of the antisense compounds described herein, as well as compounds having non-identical bases to the antisense compounds provided herein, are also contemplated. Non-identical bases may be dispersed throughout the antisense compound or adjacent to each other. The percent identity of an antisense compound is calculated according to the number of bases having identical base pairs to the sequences being compared.

In certain embodiments, the antisense compound, or portion thereof, is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least one or more of an antisense compound or SEQ ID NO, or portion thereof disclosed herein. 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical.

transform

Nucleosides are base-sugar combinations. The nucleobase (also known as a base) portion of a nucleoside is usually a heterocyclic base moiety. A nucleotide is a nucleoside further comprising a phosphate group covalently linked to the sugar moiety of the nucleoside. For nucleosides comprising pentofuranosyl sugars, the phosphate group may be linked to the 2', 3' or 5' hydroxyl moiety of the sugar. Oligonucleotides are formed through covalent linkage of adjacent nucleosides to each other to form linear polymeric oligonucleotides. Within an oligonucleotide structure, a phosphate group is usually considered to form the internucleoside linkage of the oligonucleotide.

Modifications to antisense compounds encompass substitutions or changes to internucleoside linkages, sugar moieties, or nucleobases. Modified antisense compounds are often compared to native forms because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for nucleic acid targets, increased stability in the presence of nucleases, or increased inhibitory activity. desirable.

Chemically modified nucleosides can also be used to increase the binding affinity of a shortened or truncated antisense oligonucleotide for its target nucleic acid. Consequently, comparable results can often be obtained with shorter antisense compounds with such chemically modified nucleosides.

Modified internucleoside linkages

"Naturally occurring internucleoside linkages" in RNA and DNA refer to 3' to 5' phosphodiester linkages. Antisense compounds having one or more modified, ie, non-naturally occurring, internucleoside linkages, exhibit desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for the target nucleic acid, and presence of nucleases. It is often chosen over antisense compounds with naturally occurring internucleoside linkages because of their increased stability.

Oligonucleotides with modified internucleoside linkages include internucleoside linkages having phosphorus atoms as well as internucleoside linkages without phosphorus atoms. Representative phosphorus-containing internucleoside linkages include, but are not limited to, phosphodiesters, phosphotriesters, methylphosphonates, phosphoramidates, and phosphorothioates. Methods for preparing phosphorus-containing and non-phosphorus-containing linkages are known.

In certain embodiments, an antisense compound targeted to an ANGPTL3 nucleic acid comprises one or more modified internucleoside linkages. In certain embodiments, the modified internucleoside linkage is a phosphorothioate linkage. In certain embodiments, each internucleoside linkage of an antisense compound is a phosphorothioate internucleoside linkage.

In certain embodiments, the oligonucleotide comprises modified internucleoside linkages arranged along the oligonucleotide or region thereof in a defined pattern or modified internucleoside linkage motif. In certain embodiments, the internucleoside linkages are arranged in a gapped motif. In such embodiments, the internucleoside linkage in each of the two wing regions is different from the internucleoside linkage in the gap region. In such embodiments, the internucleoside linkage in the wing region is a phosphodiester and the internucleoside linkage in the gap is a phosphorothioate. Nucleoside motifs are selected independently, so such an oligonucleotide with a gapped internucleoside linkage may not have a gapped nucleoside, and if it does not have a gapped nucleoside, the wing and gap lengths may or may not be the same.

In certain embodiments, the oligonucleotide comprises regions with alternating internucleoside linkage motifs. In certain embodiments, oligonucleotides of the invention comprise regions of uniformly modified internucleoside linkages. In certain such embodiments, the oligonucleotide comprises regions uniformly linked by phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotides are homogeneously linked by phosphorothioates. In certain embodiments, each internucleoside linkage of the oligonucleotide is selected from a phosphodiester and a phosphorothioate. In certain embodiments, each internucleoside linkage of the oligonucleotide is selected from phosphodiester and phosphorothioate and at least one internucleoside linkage is phosphorothioate.

In certain embodiments, the oligonucleotide comprises at least 6 phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least 8 phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least 10 phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least 6 consecutive phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least 8 consecutive phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least 10 consecutive phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least a block of at least one 12 consecutive phosphorothioate internucleoside linkages. In certain such embodiments, at least one such block is located at the 3' end of the oligonucleotide. In certain such embodiments, at least one such block is located within three nucleosides of the 3' end of the oligonucleotide.

In certain embodiments, the oligonucleotide comprises one or more methylphosphonate linkages. In certain embodiments, an oligonucleotide having a gapmer nucleoside motif comprises a linking motif comprising all phosphorothioate linkages except one or two methylphosphonate linkages. In certain embodiments, one methylphosphonate linkage is in the central gap of an oligonucleotide having a gapmer nucleoside motif.

In certain embodiments, it is desirable to arrange the number of phosphorothioate internucleoside linkages and phosphodiester internucleoside linkages to maintain nuclease resistance. In certain embodiments, it is desirable to arrange the number and position of phosphorothioate internucleoside linkages and the number and position of phosphodiester internucleoside linkages to maintain nuclease resistance. In certain embodiments, the number of phosphorothioate internucleoside linkages may decrease and the number of phosphodiester internucleoside linkages may increase. In certain embodiments, the number of phosphorothioate internucleoside linkages may decrease and the number of phosphodiester internucleoside linkages may increase while still maintaining nuclease resistance. In certain embodiments, it is desirable to reduce the number of phosphorothioate internucleoside linkages while maintaining nuclease resistance. In certain embodiments, it is desirable to increase the number of phosphodiester internucleoside linkages in order to maintain nuclease resistance.

modified sugar moieties

Antisense compounds may optionally contain one or more nucleosides, wherein the sugar group has been modified. Such sugar modified nucleosides may confer enhanced nuclease stability, increased binding affinity, or some other beneficial biological property to antisense compounds. In certain embodiments, the nucleoside comprises a chemically modified ribofuranose ring moiety. Examples of chemically modified ribofuranose rings include, but are not limited to, the addition of substituent groups (5' and 2' substituent groups, of non-geminal ring atoms to form bicyclic nucleic acids (BNA)). bridging, including replacement of S, N(R), or C(R ₁ )(R ₂ ) of a ribosyl ring oxygen atom) (each of R, R ₁ and R ₂ is independently H, C ₁ -C ₁₂ alkyl or protecting groups) and combinations thereof. Examples of chemically modified sugars include: 2'-F-5'-methyl substituted nucleosides (see, PCT International Application WO 2008/101157, other disclosed 5', 2'-bis substituted nucleosides) Published 8/21/08 for seed), or replacement of a ribosyl ring oxygen atom by S by further substitution at the 2'-position (see, Published US Patent Application US2005/0130923, Jun. 16, 2005) ), or alternatively, 5'-substitution of BNA (see, PCT International Application WO 2007/134181, published in 11/22/07, LNA is, for example, 5'-methyl or 5'-vinyl substituted by groups).

Examples of nucleosides with modified sugar moieties are 5'-vinyl, 5'-methyl (R or S), 4'-S, 2'-F, 2'-OCH ₃ , 2'-OCH ₂ CH ₃ , 2′-OCH ₂ CH ₂ F and 2′-O(CH ₂ ) ₂ OCH ₃ substituent groups. Substituents at the 2' position are allyl, amino, azido, thio, O-allyl, OC ₁ -C ₁₀ alkyl, OCF ₃ , OCH ₂ F, O(CH ₂ ) ₂ SCH ₃ , O(CH ₂ ) ₂ - ON(R _m )(R _n ), O-CH ₂ -C(=O)-N(R _m )(R _n ), and O-CH ₂ -C(=O)-N(R ₁ )-( CH ₂ ) ₂ -N(R _m )(R _n ), wherein each R ₁ , R _m and R _n is independently H or substituted or unsubstituted C ₁ -C ₁₀ alkyl .

As used herein, "bicyclic nucleoside" means a modified nucleoside comprising a bicyclic sugar moiety. Examples of bicyclic nucleic acids (BNAs) include, but are not limited to, nucleosides comprising a bridge between 4' and 2' ribosyl ring atoms. In certain embodiments, antisense compounds provided herein comprise one or more BNA nucleosides, wherein the bridge comprises one of the formulas: 4'-(CH ₂ )-0-2'(LNA);4'-(CH ₂ )-S-2'; 4′-(CH ₂ ) ₂ —O-2′ (ENA); 4'-CH(CH ₃ )--O-2' and 4'-CH(CH ₂ OCH ₃ )--O-2' (and analogs thereof, see US Pat. No. 7,399,845, issued Jul. 15, 2008) ); 4′-C(CH ₃ )(CH ₃ )——O-2′ (and analogs thereof, see PCT/US2008/068922, published Jan. 8, 2009 as WO/2009/006478); 4′-CH ₂ —N(OCH ₃ )-2′ (and analogs thereof, see PCT/US2008/064591, published December 11, 2008 as WO/2008/150729); 4'-CH ₂ -ON(CH ₃ )-2' (see International Application Publication US2004-0171570, published September 23, 2004); 4'-CH2-N(R)-O-2', wherein R is H, C ₁ -C ₁₂ alkyl, or a protecting group (see US Pat. No. 7,427,672, issued Sep. 23, 2008); 4'-CH ₂ -C-(H)(CH ₃ )-2' (Zhou et al., J. Org. Chem., 2009, 74 , 118-134); and 4'-CH ₂ -C-(=CH ₂ )-2' (and analogs thereof, see PCT/US2008/066154, published December 8, 2008 as WO 2008/154401).

Additional reports regarding bicyclic nucleotides can be found in the following publications (see, e.g., Srivastava et al., J. Am. Chem. Soc ., 2007, 129(26) 8362-8379; Frieden et al. , Nucleic Acids Research, 2003, 21 , 6365-6372; Elayadi et al., Curr. Opinion Invens. Drugs, 2001, 2 , 558-561; Braasch et al., Chem. Biol., 2001, 8 , 1-7 Orum et al., Curr. Opinion Mol. Ther., 2001, 3 , 239-243; Wahlestedt et al., Proc. Natl. Acad. Sci. USA , 2000, 97 , 5633-5638; Singh et al., Chem. Commun. , 1998, 4 , 455-456; Koshkin et al., Tetrahedron , 1998, 54 , 3607-3630; Kumar et al., Bioorg. Med. Chem. Lett ., 1998, 8 , 2219-2222; Singh et al., J. Org. Chem ., 1998, 63 , 10035-10039; US Patent Nos: 7,741,457; 7,399,845; 7,053,207; 7,034,133; 6,794,499; 6,770,748; 6,670,461; 6,525,0039618; 6,268,490; US2007-0287831;US2004-0171570;US Patent Application Serial Numbers: 61/097,787; 61/026,995; and International Applications: WO 2009/006478; WO 2008/154401; WO 2008/150729; WO 2009/100320; WO 2011/ 017521; WO 2009/067647; WO 2010/036698; WO 2007/134181; WO 2005/021570; WO 2004/106356; WO 99/14226. Each of the above bicyclic nucleosides can be prepared to have one or more stereochemical sugar configurations comprising, for example, α-L-ribofuranose and β-D-ribofuranose (see PCT International Application PCT/DK98/00393) , published Mar. 25, 1999 as WO 99/14226).

As used herein, "monocyclic nucleoside" means a nucleoside comprising a modified sugar moiety that is not a bicyclic sugar moiety. In certain embodiments, the sugar moiety, or sugar moiety analog, of a nucleoside may be modified or substituted at any position.

As used herein, "4'-2' bicyclic nucleoside" or "4' to 2' bicyclic nucleoside" is a fura comprising a bridge connecting the 2' carbon atom and the 4' carbon atom of a sugar ring. It refers to a bicyclic nucleoside containing a north ring.

In certain embodiments, the bicyclic sugar moiety of a BNA nucleoside includes, but is not limited to, compounds having at least one bridge between the 4' and 2' positions of a carbon atom of a pentofuranosyl sugar moiety, wherein Such bridges are independently -[C(R _a )(R _b )] _n -, -C(R _a )=C(R _b )-, -C(R _a )=N-, -C( =NR _a )-, -C(=O)-, -C(=S)-, -O-, -Si(R _a ) ₂ -, -S(=O) _x -, and -N(R _{a )} ) - comprising 1 or 2 to 4 linked groups independently selected from; where x is 0, 1, or 2; n is 1, 2, 3, or 4; each R _a and R _b is, independently, H, protecting group, hydroxyl, C ₁ -C ₁₂ alkyl, substituted C ₁ -C ₁₂ alkyl, C ₂ -C ₁₂ alkenyl, substituted C ₂ -C ₁₂ alkenyl, C ₂ -C ₁₂ alkynyl, substituted C ₂ -C ₁₂ alkynyl, C ₅ -C ₂₀ aryl, substituted C ₅ -C ₂₀ aryl, heterocycle radical, substituted heterocycle radical, heteroaryl, substituted heteroaryl, C ₅ -C ₇ cycloaliphatic radical, substituted C ₅ -C ₇ cycloaliphatic radical, halogen, OJ ₁ , NJ ₁ J ₂ , SJ ₁ , N ₃ , COOJ ₁ , acyl (C(=O )-H), substituted acyl, CN, sulfonyl (S(=O) ₂ -J ₁ ), or sulfoxyl (S(=O)-J ₁ );

and J ₁ and each J ₂ is independently H, C ₁ -C ₁₂ alkyl, substituted C ₁ -C ₁₂ alkyl, C ₂ -C ₁₂ alkenyl, substituted C ₂ -C ₁₂ alkenyl, C ₂ -C ₁₂ alkynyl , substituted C ₂ -C ₁₂ alkynyl, C ₅ -C ₂₀ aryl, substituted C ₅ -C ₂₀ aryl, acyl (C(=O)-H), substituted acyl, heterocyclic radical, substituted heterocycle radical, C ₁ -C ₁₂ aminoalkyl, substituted C ₁ -C ₁₂ aminoalkyl or a protecting group.

In certain embodiments, the bridging of the bicyclic sugar moiety is: -[C(R _a )(R _b )] _n -, -[C(R _a )(R _b )] _n -O-, -C(R _a R _b )-N(R)-O- or -C(R _a R _b )-ON(R)-. In certain embodiments, the bridge is 4'-CH ₂ -2', 4'-(CH ₂ ) ₂ -2', 4'-(CH ₂ ) ₃ -2', 4'-CH ₂ -O-2' , 4'-(CH ₂ ) ₂ -O-2', 4'-CH ₂ -ON(R)-2' and 4'-CH ₂ -N(R)-O-2'-, wherein each R is independently H, a protecting group, or C ₁ -C ₁₂ alkyl.

In certain embodiments, bicyclic nucleosides are further defined by the isomeric configuration. For example, a nucleoside comprising a 4'-(CH ₂ )-O-2' bridge may be in the α-L configuration or the β-D configuration. Previously, α-L-methyleneoxy (4′-CH ₂ —O-2′) BNA′ was incorporated into antisense oligonucleotides that showed antisense activity (Frieden et al., Nucleic Acids Research, 2003, 21 , 6365- 6372).

In certain embodiments, bicyclic nucleosides include those with 4′ to 2′ bridges, including but not limited to α-L-4′-(CH ₂ )-O-2′, β-D-4 '-CH ₂ -O-2', 4'-(CH ₂ ) ₂ -O-2', 4'-CH ₂ -ON(R)-2', 4'-CH ₂ -N(R)-O -2', 4'-CH(CH ₃ )-O-2', 4'-CH ₂ -S-2', 4'-CH ₂ -N(R)-2', 4'-CH ₂ -CH (CH ₃ )-2′, and 4′-(CH ₂ ) ₃ -2′, wherein R is H, a protecting group, or C ₁ -C ₁₂ alkyl.

In certain embodiments, the bicyclic nucleoside has the formula:

In the above formula,

Bx is a heterocyclic base moiety;

-Q _a -Q _b -Q _c - is -CH ₂ -N(R _c )-CH ₂ -, -C(=O)-N(R _c )-CH ₂ -, -CH ₂ -ON(R _c )-, -CH ₂ -N(R _c )-O- or -N(R _c )-O-CH ₂ ;

R _c is a C ₁ -C ₁₂ alkyl or amino protecting group; and

each of T _a and T _b is, independently, H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety, or a covalent attachment to a support medium;

In certain embodiments, the bicyclic nucleoside has the formula:

In the above formula,

Bx is a heterocyclic base moiety;

each of T _a and T _b is, independently, H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety, or a covalent attachment to a support medium;

Z _a is C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, substituted C ₁ -C ₆ alkyl, substituted C ₂ -C ₆ alkenyl, substituted C ₂ - C ₆ alkynyl, acyl, substituted acyl, substituted amide, thiol, or substituted thiol.

In one embodiment, each substituted group is, independently, halogen, oxo, hydroxyl, OJ _c , NJ _c J _d , SJ _c , N ₃ , OC(=X)J _c , and NJ _e C (= X) single or multiple substituted with a substituent group independently selected from _{NJ c} J _{d , wherein each J c} , J _d and J _e is, independently, H, C ₁ -C ₆ alkyl, or substituted C ₁ - C ₆ alkyl and X is O or NJ _c .

In certain embodiments, the bicyclic nucleoside has the formula:

In the above formula,

Bx is a heterocyclic base moiety;

each of T _a and T _b is, independently, H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety, or a covalent attachment to a support medium;

Z _b is C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, substituted C ₁ -C ₆ alkyl, substituted C ₂ -C ₆ alkenyl, substituted C ₂ - C ₆ alkynyl, or substituted acyl(C(=O)—).

In certain embodiments, the bicyclic nucleoside has the formula:

In the above formula,

Bx is a heterocyclic base moiety;

each of T _a and T _b is, independently, H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety, or a covalent attachment to a support medium;

R _d is C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, substituted C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, or substituted C ₂ -C ₆ alkynyl;

each q _a , q _b , q _c and q _d is independently H, halogen, C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, substituted C ₂ - C ₆ alkenyl, C ₂ -C ₆ alkynyl or substituted C ₂ -C ₆ alkynyl, C ₁ -C ₆ alkoxyl, substituted C ₁ -C ₆ alkoxyl, acyl, substituted acyl, C ₁ - C ₆ aminoalkyl or substituted C ₁ -C ₆ aminoalkyl;

In certain embodiments, the bicyclic nucleoside has the formula:

In the above formula,

Bx is a heterocyclic base moiety;

each of T _a and T _b is, independently, H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety, or a covalent attachment to a support medium;

q _a , q _{b ,} q _e and q _f each is, independently, hydrogen, halogen, C ₁ -C ₁₂ alkyl, substituted C ₁ -C ₁₂ alkyl, C ₂ -C ₁₂ alkenyl, substituted C ₂ -C ₁₂ alkenyl, C ₂ -C ₁₂ alkyl nyl, substituted C ₂ -C ₁₂ alkynyl, C ₁ -C ₁₂ alkoxy, substituted C ₁ -C ₁₂ alkoxy, OJ _j , SJ _j , SOJ _j , SO ₂ J _j , NJ _j J _k , N ₃ , CN, C(=O)OJ _j , C(=O)NJ _j J _k , C(=O)J _j , OC(=O)-NJ _j J _k , N(H)C(=NH)NJ _j J _k , N(H)C(=O)-NJ _j J _k or N(H)C(=S)NJ _j J _k ;

or q _e and q _f together =C(q _g )(q _h );

each of q _g and q _h is, independently, H, halogen, C ₁ -C ₁₂ alkyl, or substituted C ₁ -C ₁₂ alkyl.

Synthesis and preparation of adenine, cytosine, guanine, 5-methyl-cytosine, thymine and uracil bicyclic nucleosides with 4'-CH _{2 -O-2' bridges, their oligomerization, and nucleic acid recognition properties are described.} (Koshkin et al., Tetrahedron , 1998, 54 , 3607-3630). The synthesis of bicyclic nucleosides is also described in WO 98/39352 and WO 99/14226.

Analogs of various bicyclic nucleosides having 4' to 2' bridging groups such as 4'-CH ₂ -O-2' and 4'-CH ₂ -S-2' have also been prepared (Kumar et al., Bioorg. Med. Chem. Lett. , 1998, 8 , 2219-2222). The preparation of oligodeoxyribonucleotide duplexes comprising bicyclic nucleosides for use as substrates for nucleic acid polymerases has also been described (Wengel et al., WO 99/14226). Moreover, the synthesis of 2'-amino-BNA, a novel conformationally restricted high-affinity oligonucleotide analog, has been described in the art (Singh et al., J. Org. Chem. , 1998, 63 , 10035- 10039). In addition, 2'-amino- and 2'-methylamino-BNA' have been prepared and the thermal stability of their duplexes with complementary RNA and DNA strands has been previously reported.

In certain embodiments, the bicyclic nucleoside has the formula:

In the above formula,

Bx is a heterocyclic base moiety;

each of T _a and T _b is, independently, H, a hydroxyl protecting group, a conjugate group, a reactive phosphorus group, a phosphorus moiety, or a covalent attachment to a support medium;

each of q _i , q _j , q _k and q _l is, independently, hydrogen, halogen, C ₁ -C ₁₂ alkyl, substituted C ₁ -C ₁₂ alkyl, C ₂ -C ₁₂ alkenyl, substituted C ₂ - C ₁₂ alkenyl, C ₂ -C ₁₂ alkynyl, substituted C ₂ -C ₁₂ alkynyl, C ₁ -C ₁₂ alkoxyl, substituted C ₁ -C ₁₂ alkoxyl, OJ _j , SJ _j , SOJ _j , SO ₂ J _j , NJ _j J _k , N ₃ , CN, C(=O)OJ _j , C(=O)NJ _j J _k , C(=O)J _j , OC(=O)-NJ _j J _k , N(H)C(=NH)NJ _j J _k , N(H)C(=O)-NJ _j J _k or N(H)C(=S)NJ _j J _k ; and

q _i and q _j or q _l and q _{k taken} together =C(q _g )(q _h ), where q _g and q _h are each independently H, halogen, C ₁ -C ₁₂ alkyl or substituted C ₁ -C ₁₂ alkyl.

One carbocyclic bicyclic nucleoside with a 4'-(CH ₂ ) ₃ -2' bridge and an alkenyl analog, a bridge 4'-CH=CH-CH ₂ -2', have been described (Frier et al. , Nucleic Acids Research , 1997, 25 ( 22 ), 4429-4443 and Albaek et al., J. Org . Chem . , 2006, 71 , 7731-7740). The synthesis and preparation of carbocyclic bicyclic nucleosides, along with their oligomerization and biochemical studies, have also been described (Srivastava et al., J. Am. Chem. Soc . 2007, 129(26) , 8362-8379). ).

In certain embodiments, the bicyclic nucleoside is, without limitation, (A) α-L-methyleneoxy (4′-CH ₂ —O-2′) BNA, (B) β-D-methyleneoxy (4′- CH ₂ -O-2′) BNA, (C) ethyleneoxy (4′-(CH ₂ ) ₂ -O-2′) BNA, (D) aminooxy (4′-CH ₂ -ON(R)-2 ') BNA, (E) oxyamino (4'-CH ₂ -N(R)-O-2') BNA, and (F) methyl(methyleneoxy) (4'-CH(CH ₃ )-O-2 ') BNA (also referred to as restricted ethyl or cEt), (G) methylene-thio (4'-CH ₂ -S-2') BNA, (H) methylene-amino (4'-CH ₂ -N(R) )-2') BNA, (I) methyl carbocyclic (4'-CH ₂ -CH(CH ₃ )-2') BNA, (J) propylene carbocyclic (4'-(CH ₂ ) ₃ -2') BNA and (K) vinyl BNA, as depicted below:

wherein Bx is a base moiety and R is, independently, H, a protecting group or C ₁ -C ₆ alkyl or C ₁ -C ₆ alkoxy.

As used herein, "modified tetrahydropyran nucleoside" or "modified THP nucleoside" refers to a 6-membered tetrahydropyran "sugar substituted to replace a pentofuranosyl residue in the normal nucleoside". " means a nucleoside with ", and may be referred to as a sugar substitute. Modified THP nucleosides include, but are not limited to, those referred to in the art as hexitol nucleic acid (HNA), anthole nucleic acid (ANA), mannitol nucleic acid (MNA) (see Leumann, Bioorg . Med . Chem ., 2002 , 10 , 841-854) or fluoro HNA (F-HNA) (which has a tetrohydropyranyl ring system as exemplified below).

In certain embodiments, the sugar surrogate is selected from the formula:

In the above formula,

Bx is a heterocyclic base moiety;

each of T ₃ and T ₄ is, independently, an internucleoside linking group linking a tetrahydropyran nucleoside analog to an oligomeric compound or one of _{T 3} and T _{4 is a tetrahydropyran nucleoside analog to an oligomeric compound or} an internucleoside linking group linking to the oligonucleotide, and the other of T ₃ and T ₄ is H, a hydroxyl protecting group, a linked conjugate group, or a 5′ or 3′-end group;

each of q ₁ , q ₂ , q ₃ , q ₄ , q ₅ , q ₆ and q ₇ is independently H, C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, substituted C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl or substituted C ₂ -C ₆ alkynyl; and

one of R ₁ and R ₂ is hydrogen and the other is halogen, substituted or unsubstituted alkoxy, NJ ₁ J ₂ , SJ ₁ , N ₃ , OC(=X)J ₁ , OC(=X)NJ ₁ J ₂ , NJ ₃ C(=X)NJ ₁ J ₂ and CN, wherein X is O, S or NJ ₁ and each J ₁ , J ₂ and J ₃ is, independently, H or C ₁ -C ₆ alkyl.

In certain embodiments, each of q ₁ , q ₂ , q ₃ , q ₄ , q ₅ , q ₆ and q ₇ is H. In certain embodiments, at least one of _{q 1} , q ₂ , q ₃ , q ₄ , q ₅ , q ₆ and q _{7 is other than H.} In certain embodiments, at least one of _{q 1} , q ₂ , q ₃ , q ₄ , q ₅ , q ₆ and q _{7 is methyl.} In certain embodiments, provided are THP nucleosides, wherein one of _{R 1} and R _{2 is F.} In certain embodiments, R ₁ is fluoro and R ₂ is H; R ₁ is methoxy and R ₂ is H, and R ₁ is methoxyethoxy and R ₂ is H.

In certain embodiments, sugar surrogates include rings having more than 5 atoms and more than 1 heteroatom. For example, their use in nucleosides and oligomeric compounds comprising morpholino modified sugar moieties has been reported (see, for example , Braasch et al ., Biochemistry , 2002, 41 , 4503-4510; and U.S. Patents 5,698,685; 5,166,315; 5,185,444; and 5,034,506). As used herein, "morpholino" means a sugar surrogate having the structure:

.

.

In certain embodiments, a morpholino can be modified by adding or altering various substituent groups from the morpholino structure. Such sugar surrogates are referred to herein as "modified morpholinos".

Combinations of modifications are also provided, but are not limited to: 2'-F-5'-methyl substituted nucleosides (see 8/21/ for other disclosed 5', 2'-bis substituted nucleosides) PCT International Application WO 2008/101157, published 08) and substitution and further substitution with S for the ribosyl ring oxygen atom in the 2'-position (see US Patent Application US2005-0130923 published on Jun. 16, 2005) ) or alternatively a 5'-substitution of a bicyclic nucleic acid (PCT International Application WO 2007/134181 published ref. 11/22/07, wherein the 4'-CH ₂ -O-2' bicyclic nucleoside is 5' at the 5' position further substituted with a '-methyl or 5'-vinyl group). The synthesis and preparation of carbocyclic bicyclic nucleosides, along with their oligomerization and biochemical studies, have also been described (see, e.g., Srivastava et al., J. Am. Chem. Soc. 2007, 129(26)). , 8362-8379).

et al., Tetrahedron Letters, 2007, 48, 3621-3623; Nauwelaerts et al., J. Am. Chem . Soc., 2007, 129(30), 9340-9348; Gu et al.,, Nucleosides , Nucleotides & Nucleic Acids, 2005, 24(5-7), 993-998; Nauwelaerts et al., Nucleic Acids Research, 2005, 33(8), 2452-2463; Robeyns et al., Acta Crystallographica , Section F: Structural Biology and Crystallization Communications, 2005, F61(6), 585-586; Gu et al., Tetrahedron, 2004, 60(9), 2111-2123; Gu et al., Oligonucleotides, 2003, 13(6), 479-489; Wang et al., J. Org . Chem ., 2003, 68, 4499-4505; Verbeure et al., Nucleic Acids Research, 2001, 29(24), 4941-4947; Wang et al., J. Org . Chem., 2001, 66, 8478-82; Wang et al., Nucleosides, Nucleotides & Nucleic Acids, 2001, 20(4-7), 785-788; Wang et al., J. Am. Chem ., 2000, 122, 8595-8602; 공개 PCT 출원, WO 06/047842; 및 공개 PCT 출원 WO 01/049687; 각각의 내용은 본원에 그 전체가 참고로 편입되어 있다). 특정 변형된 사이클로헥세닐 뉴클레오시드는 하기 화학식 X를 가진다. In certain embodiments, antisense compounds comprise one or more modified cyclohexenyl nucleosides, which are nucleosides having a 6-membered cyclohexenyl in place of the pentofuranosyl present in the naturally occurring nucleoside. Modified cyclohexenyl nucleosides include, but are not limited to, those described in the art in the following (eg, generally reserved, see Published PCT Application WO 2010/036696, published Apr. 10, 2010) , Robeyns et al., J. Modified cyclohexynyl nucleosides include, but are not limited to, those described in the art: (eg, generally retained, see Published PCT Application WO 2010/ 036696 (published April 10, 2010), Robeyns et al., J. Am. Chem . Soc ., 2008, 130(6) , 1979-1984;

et al., Tetrahedron Letters, 2007, 48 , 3621-3623; Nauwelaerts et al., J. Am. Chem . Soc ., 2007, 129(30) , 9340-9348; Gu et al., , Nucleosides , Nucleotides & Nucleic Acids , 2005, 24(5-7) , 993-998; Nauwelaerts et al., Nucleic Acids Research, 2005, 33(8) , 2452-2463; Robeyns et al., Acta Crystallographica , Section F: Structural Biology and Crystallization Communications, 2005, F61(6) , 585-586; Gu et al., Tetrahedron , 2004, 60(9) , 2111-2123; Gu et al., Oligonucleotides , 2003, 13(6) , 479-489; Wang et al., J. Org . Chem ., 2003, 68 , 4499-4505; Verbeure et al., Nucleic Acids Research , 2001, 29(24) , 4941-4947; Wang et al., J. Org . Chem., 2001, 66 , 8478-82; Wang et al. , Nucleosides, Nucleotides & Nucleic Acids , 2001, 20(4-7) , 785-788; Wang et al., J. Am. Chem ., 2000, 122 , 8595-8602; published PCT applications, WO 06/047842; and published PCT applications WO 01/049687; The contents of each are incorporated herein by reference in their entirety). Certain modified cyclohexenyl nucleosides have the formula (X):

wherein, independently for each of the at least one cyclohexenyl nucleoside analog of formula X:

Bx is a heterocyclic base moiety.

each of T ₃ and T ₄ is, independently, an internucleoside linking group linking the cyclohexenyl nucleoside analog to its antisense compound or _{one of T 3} and T ₄ is a tetrahydropyran nucleoside analog to the antisense compound an internucleoside linking group linking to, and the other of T ₃ and T ₄ is H, a hydroxyl protecting group, a linked conjugate group, or a 5' or 3'-terminal group;

q ₁ , q ₂ , q ₃ , q ₄ , q ₅ , q ₆ , q ₇ , q ₈ and q ₉ are each, independently, H, C ₁ -C ₆ alkyl, substituted C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, substituted C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, substituted C ₂ -C ₆ alkynyl or other sugar substituent group.

Many other monocyclic, bicyclic and tricyclic ring systems are known in the art and are suitable as sugar substitutes that can be used to modify nucleosides for incorporation into oligomeric compounds as provided herein (e.g. See, for example, the following review article: Leumann, Christian J. Bioorg . & Med . Chem . , 2002, 10 , 841-854). Such ring systems may undergo various additional substitutions to further enhance activity.

As used herein, "2'-modified sugar" refers to a furanosyl sugar modified at the 2' position. In certain embodiments, such modifications are selected from substituents including, but not limited to, substituted and unsubstituted alkoxy, substituted and unsubstituted thioalkyl, substituted and unsubstituted amino alkyl, substituted and unsubstituted Halides including, but not limited to, substituted alkyl, substituted and unsubstituted allyl, and substituted and unsubstituted alkynyl. In certain embodiments, the 2' modification is selected from substituents including, but not limited to: O[(CH ₂ ) _n O] _m CH ₃ , O(CH ₂ ) _n NH ₂ , O(CH ₂ ) _n CH ₃ , O(CH ₂ ) _n F, O(CH ₂ ) _n ONH ₂ , OCH ₂ C(=O)N(H)CH _{3 ,} and O(CH ₂ ) _n ON[(CH ₂ ) _n CH ₃ ] ₂ , wherein n and m are from 1 to about 10. Other 2'-substituent groups may also be selected from: C ₁ -C ₁₂ alkyl; substituted alkyl; alkenyl; alkynyl; alkaryl; aralkyl; O-alkaryl or O-aralkyl; SH, SCH ₃ , OCN, Cl, Br, CN, F, CF ₃ , OCF ₃ , SOCH ₃ , SO ₂ CH ₃ , ONO ₂ , NO ₂ , N ₃ , NH ₂ ; heterocycloalkyl; heterocycloalkaryl; aminoalkylamino; polyalkylamino; substituted silyl; RNA cleavage group; reporter group; insert; groups that improve pharmacodynamic properties; and groups that improve the pharmacokinetic properties of antisense compounds, and other substituents having similar properties. In certain embodiments, the modified nucleoside comprises a 2'-MOE side chain (Baker et al ., J. Biol . Chem . , 1997, 272 , 11944-12000). Such 2'-MOE substitutions have improved binding affinity compared to unmodified nucleosides and other modified nucleosides, such as 2'-O-methyl, O-propyl, and O-aminopropyl. was described as Oligonucleotides with 2'-MOE substitutions have also been shown to be antisense inhibitors of gene expression with promising properties for in vivo use (cf. Martin, Helv. Chim . Acta , 1995, 78 , 486-504; Altmann et al ., Chimia , 1996, 50 , 168-176; Altmann et al ., Biochem . Soc . Trans ., 1996, 24 , 630-637; and Altmann et al ., Nucleosides Nucleotides , 1997, 16 , 917-926).

As used herein, "2'-modified" or "2'-substituted" refers to a nucleoside comprising a sugar comprising a substituent at the 2' position other than H or OH. 2'-modified nucleosides include, but are not limited to, bicyclic nucleosides, wherein the bridge connecting two carbon atoms of the sugar ring is the 2' carbon of the sugar ring and the other carbon; and a nucleoside with a non-bridging 2' substituent such as allyl, amino, azido, thio, O-allyl, OC ₁ -C ₁₀ alkyl, -OCF ₃ , O-(CH ₂ ) ₂ -O-CH ₃ , 2'-O(CH ₂ ) ₂ SCH ₃ , O-(CH ₂ ) ₂ -ON(R _m )(R _n ), or O-CH ₂ -C(=O)-N(R _m )(R _{n )} ), wherein each of R _m and R _n is, independently, H or substituted or unsubstituted C ₁ -C ₁₀ alkyl. 2'-modified nucleosides may further comprise other modifications, for example at other positions of the sugar and/or at the nucleobase.

As used herein, “2′-F” refers to a nucleoside comprising a sugar comprising a fluoro group at the 2′ position of the sugar ring.

As used herein, "2'-OMe" or "2'-OCH ₃ " or "2'-O-methyl" or "2'-methoxy" each represents an -OCH ₃ group at the 2' position of the sugar ring. refers to a nucleoside comprising a sugar comprising

As used herein, “MOE” or “2′-MOE” or “2′-OCH ₂ CH ₂ OCH ₃ ” or “2′-O-methoxyethyl” respectively is -OCH at the 2′ position of the sugar ring refers to a nucleoside comprising a sugar comprising a ₂ CH ₂ OCH _{3 group.}

Methods for preparing modified saccharides are well known to those skilled in the art. Some representative US patents that teach the preparation of such modified sugars include, without limitation, US Pat. Nos. 4,981,957; 5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785; 5,519,134; 5,567,811; 5,576,427; 5,591,722; 5,597,909; 5,610,300; 5,627,053; 5,639,873; 5,646,265; 5,670,633; 5,700,920; 5,792,847 and 6,600,032 and International Application PCT/US 2005/019219, filed on June 2, 2005 and published as WO 2005/121371 on December 22, 2005, certain of which are incorporated herein by reference. are commonly owned, each of which is incorporated herein by reference in its entirety.

As used herein, "oligonucleotide" refers to a compound comprising a plurality of linked nucleosides. In certain embodiments, one or more of the plurality of nucleosides is modified. In certain embodiments, the oligonucleotide comprises one or more ribonucleosides (RNA) and/or deoxyribonucleosides (DNA).

In nucleotides with modified sugar moieties, the nucleobase moiety (native, modified, or a combination thereof) is maintained for hybridization with an appropriate nucleic acid target.

In certain embodiments, antisense compounds include nucleosides having one or more modified sugar moieties. In certain embodiments, the modified sugar moiety is 2'-MOE. In certain embodiments, the 2'-MOE modified nucleoside is arranged in a gapmer motif. In certain embodiments, the modified sugar moiety is _{a bicyclic nucleoside having a (4′-CH(CH 3} )-O-2′) bridging group. In certain embodiments, the (4′-CH(CH ₃ )-O-2′) modified nucleoside is arranged across the wings of the gapmer motif.

deformed nucleobase

Nucleobase (or base) modifications or substitutions are structurally distinguishable and functionally interchangeable with naturally occurring or synthetic unmodified nucleobases. Both natural and modified nucleobases can participate in hydrogen bonding. Such nucleobase modifications may confer nuclease stability, binding affinity or some other beneficial biological property to the antisense compound. Modified nucleobases include synthetic and natural nucleobases such as, for example, 5-methylcytosine (5-me-C). Any nucleobase substitution, including 5-methylcytosine substitution, is particularly useful for increasing the binding affinity of an antisense compound to a target nucleic acid. For example, 5-methylcytosine substitution has been shown to increase nucleic acid duplex stability up to 0.6-1.2 °C (Sanghvi, YS, Crooke, ST and Lebleu, B., eds., Antisense Research and Applications, CRC Press, Boca). Raton, 1993, pp. 276-278).

Additional modified nucleobases include: 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and adenine and guanine, other alkyl derivatives of 2-propyl and adenine and guanine, 2 -thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl (-C≡C-CH3) other alkyl derivatives of uracil and cytosine and pyrimidine base, 6-azouracil; Cytosine and other alkynyl derivatives of thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenine and guanine, 5-halo especially 5-bromo, 5-trifluoromethyl and other 5-substituted uracil and cytosine, 7-methylguanine and 7-methyladenine, 2-F-adenine, 2-amino-adenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine and 3-deazaguanine and 3-deazaadenine.

Heterocyclic base moieties in which a purine or pyrimidine base is replaced by another heterocycle may also be included, such as 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine and 2-pyridone. Nucleobases particularly useful for increasing the binding affinity of antisense compounds include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and O-6 substituted purines, which are 2 aminopropyladenine , 5-propynyluracil and 5-propynylcytosine.

In certain embodiments, an antisense compound targeted to an ANGPTL3 nucleic acid comprises one or more modified nucleobases. In certain embodiments, a shortened or gap-widened antisense oligonucleotide targeted to an ANGPTL3 nucleic acid comprises one or more modified nucleobases. In certain embodiments, the modified nucleobase is 5-methylcytosine. In certain embodiments, each cytosine is 5-methylcytosine.

pharmaceutical composition to formulate Compositions and methods for

Antisense oligonucleotides may be admixed with pharmaceutically acceptable active or inactive substances for the manufacture of pharmaceutical compositions or formulations. Compositions and methods of formulation of pharmaceutical compositions depend on a number of criteria, including, but not limited to, route of administration, degree of disease, or dosage to be administered.

Antisense compounds targeted to ANGPTL3 nucleic acids can be used in pharmaceutical compositions by combining the antisense compound with a suitable pharmaceutically acceptable diluent or carrier. Pharmaceutically acceptable diluents include phosphate-buffered saline (PBS). PBS is a suitable diluent for use in compositions to be delivered parenterally. Accordingly, in one embodiment, a pharmaceutical composition comprising an antisense compound targeted to an ANGPTL3 nucleic acid and a pharmaceutically acceptable diluent is used in the methods described herein. In certain embodiments, the pharmaceutically acceptable diluent is PBS. In certain embodiments, the antisense compound is an antisense oligonucleotide.

A pharmaceutical composition comprising an antisense compound may contain any pharmaceutically acceptable salt, ester, or salt of such an ester, or (directly or indirectly) a biologically active metabolite or when administered to an animal, including a human. any other oligonucleotide capable of providing a residue thereof. Thus, for example, the present disclosure also relates to pharmaceutically acceptable salts, prodrugs, pharmaceutically acceptable salts of such prodrugs, and other bioisomers of antisense compounds. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.

A prodrug may comprise incorporation of an additional nucleoside at one or both termini of the antisense compound that is cleaved by an endogenous nuclease to form an active antisense compound.

conjugated antisense compound

The antisense compound may be covalently linked to one or more moieties or conjugates that enhance the activity, cellular distribution or cellular uptake of the resulting antisense oligonucleotide. Typical conjugate groups include cholesterol moieties and lipid moieties. Additional conjugate groups include carbohydrates, phospholipids, biotin, phenazine, folate, phenanthridine, anthraquinone, acridine, fluorescein, rhodamine, coumarin, and dyes.

Antisense compounds may also be modified to have one or more stabilizing groups generally attached to one or both termini of the antisense compound to improve properties such as, for example, nuclease stability. The stabilizing group includes a cap structure. These terminal modifications protect antisense compounds with terminal nucleic acids from exonuclease degradation and may aid in delivery and/or localization within cells. The cap may be at the 5′-end (5′-cap), or at the 3′-end (3′-cap), or at both ends. Cap structures are well known in the art and include, for example, inverted deoxy abasic caps. Additional 3' and 5'-stabilizing groups that can be used to cap one or both ends of a cap antisense compound to confer nuclease stability are described in WO 03/004602 published Jan. 16, 2003. including those disclosed.

In certain embodiments, the present disclosure provides a conjugated antisense compound represented by the formula:

In the above formula,

A is an antisense oligonucleotide;

B is a cleavable moiety;

C is a conjugate linker

D is a branching group

each E is a linking group;

each F is a ligand; and

q is an integer from 1 to 5;

In certain embodiments, conjugated antisense compounds are provided having the structure:

.

.

In certain embodiments, conjugated antisense compounds are provided having the structure:

.

.

In certain embodiments, conjugated antisense compounds are provided having the structure:

.

.

The present disclosure provides the following non-limiting numbered embodiments:

In the above formula,

T ₂ is a nucleoside, nucleotide, monomer subunit, or oligomeric compound.

In embodiments having more than one (eg, more than one "m" or "n") of a particular variable, unless otherwise indicated, each such particular variable is independently selected. Thus, for structures with n greater than 1, each n is selected independently, which may or may not be the same as each other.

i. Specific cutting possible moiety

In certain embodiments, the cleavable moiety is a cleavable bond. In certain embodiments, the cleavable moiety comprises a cleavable bond. In certain embodiments, the conjugate group comprises a cleavable moiety. In certain such embodiments, the cleavable moiety is attached to the antisense oligonucleotide. In certain such embodiments, the cleavable moiety is directly attached to the cell-targeting moiety. In certain such embodiments, the cleavable moiety is attached to the conjugate linker. In certain embodiments, the cleavable moiety comprises a phosphate or phosphodiester. In certain embodiments, the cleavable moiety is a cleavable nucleoside or nucleoside analog. In certain embodiments, the nucleoside or nucleoside analog comprises an optionally protected heterocyclic base selected from a purine, a substituted purine, a pyrimidine or a substituted pyrimidine. In certain embodiments, the cleavable moiety is uracil, thymine, cytosine, 4-N-benzoylcytosine, 5-methylcytosine, 4-N-benzoyl-5-methylcytosine, adenine, 6-N-benzoyladenine, guanine and is a nucleoside comprising an optionally protected heterocyclic base selected from 2-N-isobutyrylguanine. In certain embodiments, the cleavable moiety is a 2'-deoxy nucleoside attached to the 3' position of the antisense oligonucleotide by a phosphodiester linkage and to the linker by a phosphodiester or phosphorothioate linkage. . In certain embodiments, the cleavable moiety is 2'-deoxy adenosine attached to the 3' position of the antisense oligonucleotide by a phosphodiester linkage and to the linker by a phosphodiester or phosphorothioate linkage. In certain embodiments, the cleavable moiety is 2'-deoxy adenosine attached to the 3' position of the antisense oligonucleotide by a phosphodiester linkage and to the linker by a phosphodiester or phosphorothioate linkage.

In certain embodiments, the cleavable moiety is attached at the 3' position of the antisense oligonucleotide. In certain embodiments, the cleavable moiety is attached at the 5' position of the antisense oligonucleotide. In certain embodiments, the cleavable moiety is attached at the 2' position of the antisense oligonucleotide. In certain embodiments, the cleavable moiety is attached to the antisense oligonucleotide by a phosphodiester linkage. In certain embodiments, the cleavable moiety is attached to the linker by a phosphodiester or phosphorothioate linkage. In certain embodiments, the cleavable moiety is attached to the linker by a phosphodiester linkage. In certain embodiments, the conjugate group does not include a cleavable moiety.

In certain embodiments, the cleavable moiety is cleaved after the complex is administered to the animal immediately after internalization by the targeted cell. The cleavable moiety within the cell is cleaved thereby releasing the active antisense oligonucleotide. Without wishing to be bound by theory, it is believed that the cleavable moiety is cleaved by one or more nucleases within the cell. In certain embodiments, one or more nucleases cleave the phosphodiester linkage between the cleavable moiety and the linker. In certain embodiments, the cleavable moiety has a structure selected from:

wherein each of Bx, Bx ₁ , Bx ₂ , and Bx ₃ is independently a heterocyclic base moiety. In certain embodiments, the cleavable moiety has a structure selected from:

i. specific linker

In certain embodiments, the conjugate group comprises a linker. In certain such embodiments, the linker is covalently bonded to the cleavable moiety. In certain such embodiments, the linker is covalently linked to the antisense oligonucleotide. In certain embodiments, the linker is covalently linked to a cell-targeting moiety. In certain embodiments, the linker further comprises a covalent bond to a solid support. In certain embodiments, the linker further comprises a covalent bond to a protein binding moiety. In certain embodiments, the linker further comprises a covalent bond to a solid support and further comprises a covalent bond to a protein binding moiety. In certain embodiments, the linker comprises multiple positions for attachment of linked ligands. In certain embodiments, the linker comprises multiple positions for attachment of a linked ligand and is not attached to a branching group. In certain embodiments, the linker further comprises one or more cleavable bonds. In certain embodiments, the conjugate group does not include a linker.

In certain embodiments, the linker comprises at least one linear group comprising a group selected from alkyl, amide, disulfide, polyethylene glycol, ether, thioether (-S-) and hydroxylamino (-ON(H)-) groups. include In certain embodiments, linear groups include groups selected from alkyl, amide and ether groups. In certain embodiments, linear groups include groups selected from alkyl and ether groups. In certain embodiments, the linear group comprises at least one phosphorus linking group. In certain embodiments, the linear group comprises at least one phosphodiester group. In certain embodiments, the linear group comprises at least one neutral linking group. In certain embodiments, the linear group is covalently linked to a cell-targeting moiety and a cleavable moiety. In certain embodiments, the linear group is covalently linked to a cell-targeting moiety and an antisense oligonucleotide. In certain embodiments, the linear group is covalently linked to a cell-targeting moiety, a cleavable moiety and a solid support. In certain embodiments, the linear group is covalently linked to a cell-targeting moiety, a cleavable moiety, a solid support, and a protein binding moiety. In certain embodiments, a linear group comprises one or more cleavable bonds.

In certain embodiments, the linker comprises a linear group covalently attached to a scaffold group. In certain embodiments, the scaffold comprises branched aliphatic groups comprising groups selected from alkyl, amide, disulfide, polyethylene glycol, ether, thioether and hydroxylamino groups. In certain embodiments, the scaffold comprises branched aliphatic groups comprising groups selected from alkyl, amide and ether groups. In certain embodiments, the scaffold comprises at least one monocyclic or polycyclic ring system. In certain embodiments, the scaffold comprises at least two monocyclic or polycyclic ring systems. In certain embodiments, the linear group is covalently bonded to a scaffold group and the scaffold group is covalently bonded to a cleavable moiety and a linker. In certain embodiments, the linear group is covalently linked to a scaffold group and the scaffold group is covalently linked to a cleavable moiety, a linker and a solid support. In certain embodiments, the linear group is covalently linked to a scaffold group and the scaffold group is covalently linked to a cleavable moiety, a linker and a protein binding moiety. In certain embodiments, the linear group is covalently linked to a scaffold group and the scaffold group is covalently linked to a cleavable moiety, a linker, a protein binding moiety and a solid support. In certain embodiments, the scaffold group comprises one or more cleavable linkages.

In certain embodiments, the linker comprises a protein binding moiety. In certain embodiments, the protein binding moiety is a lipid including, but not limited to, cholesterol, cholic acid, adamantane acetic acid, 1-pyrene butyric acid, dihydrotestosterone, 1,3-bis-O(hexadecyl)glycerol. , geranyloxyhexyl group, hexadecylglycerol, dragonfly, menthol, 1,3-propanediol, heptadecyl group, palmitic acid, myristic acid, O3-(oleoyl) lithocholic acid, O3-(oleoyl) cholenic acid, dimethoxytrityl, or phenoxazine), vitamins (eg, folate, vitamin A, vitamin E, biotin, pyridoxal), peptides, carbohydrates (eg, monosaccharides, disaccharides, trisaccharides) Rides, tetrasaccharides, oligosaccharides, polysaccharides), endosomal components, steroids (eg ubaol, hexigenin, diosgenin), terpenes (eg triterpenes, eg sarsasapo) genin, friedelin, epipriedelanol derived lithocholic acid), or cationic lipids. In certain embodiments, the protein binding moiety is a C16 to C22 long chain saturated or unsaturated fatty acid, cholesterol, cholic acid, vitamin E, adamantane or 1-pentafluoropropyl.

In certain embodiments, the linker has a structure selected from:

wherein each n is, independently, 1 to 20; and p is 1 to 6.

In certain embodiments, the linker has a structure selected from:

In the above formula, each n is independently 1 to 20.

In certain embodiments, the linker has a structure selected from:

In the above formula, n is 1 to 20.

In certain embodiments, the linker has a structure selected from:

wherein each L is, independently, a phosphorus linking group or a neutral linking group; and

each n is, independently, 1-20.

In certain embodiments, the linker has a structure selected from:

.

.

In certain embodiments, the linker has a structure selected from:

.

.

In certain embodiments, the linker has a structure selected from:

.

.

In certain embodiments, the linker has a structure selected from:

In the above formula, n is 1 to 20.

In certain embodiments, the linker has a structure selected from:

.

.

In certain embodiments, the linker has a structure selected from:

.

.

In certain embodiments, the linker has a structure selected from:

.

.

In certain embodiments, the conjugate linker has the structure:

.

.

In certain embodiments, the conjugate linker has the structure:

.

.

In certain embodiments, the linker has a structure selected from:

.

.

In certain embodiments, the linker has a structure selected from:

;

;

wherein each n is independently 0, 1, 2, 3, 4, 5, 6, or 7.

ii. specific cell- targeting moiety

In certain embodiments, the conjugate group comprises a cell-targeting moiety. Certain such cell-targeting moieties increase the cellular uptake of antisense compounds. In certain embodiments, a cell-targeting moiety comprises a branching group, one or more linking groups, and one or more ligands. In certain embodiments, a cell-targeting moiety comprises a branching group, one or more linking groups, one or more ligands, and one or more cleavable bonds.

One. specific branching group

In certain embodiments, the conjugate group comprises a targeting moiety comprising a branching group and at least two linked ligands. In certain embodiments, the branching group attaches a conjugate linker. In certain embodiments, the branching group attaches a cleavable moiety. In certain embodiments, the branching group attaches an antisense oligonucleotide. In certain embodiments, the branching group is covalently attached to the linker and to each linked ligand. In certain embodiments, branching groups include branched aliphatic groups including groups selected from alkyl, amide, disulfide, polyethylene glycol, ether, thioether and hydroxylamino groups. In certain embodiments, branching groups include groups selected from alkyl, amide and ether groups. In certain embodiments, branching groups include groups selected from alkyl and ether groups. In certain embodiments, branching groups include monocyclic or polycyclic ring systems. In certain embodiments, the branching group comprises one or more cleavable bonds. In certain embodiments, the conjugate group does not include a branching group.

In certain embodiments, the branching group has a structure selected from:

.

.

In the above formula, each n is independently 1 to 20.

j is 1 to 3; and

m is 2 to 6.

In certain embodiments, the branching group has a structure selected from:

wherein each n is independently 1 to 20; and

m is 2 to 6.

In certain embodiments, the branching group has a structure selected from:

.

.

In certain embodiments, the branching group has a structure selected from:

wherein each A ₁ is independently O, S, C=O or NH; and

each n is, independently, 1-20.

In certain embodiments, the branching group has a structure selected from:

wherein each A ₁ is independently O, S, C=O or NH; and

each n is, independently, 1-20.

In certain embodiments, the branching group has a structure selected from:

wherein A ₁ is O, S, C=O or NH; and

each n is, independently, 1-20.

In certain embodiments, the branching group has a structure selected from:

.

.

In certain embodiments, the branching group has a structure selected from:

.

.

In certain embodiments, the branching group has a structure selected from:

.

.

2. specific coupler

In certain embodiments, the conjugate group comprises one or more linking groups covalently bonded to the branching group. In certain embodiments, the conjugate group comprises one or more linking groups covalently bonded to the linking group. In certain embodiments, each linking group is a linear aliphatic group comprising one or more groups selected from alkyl, ether, thioether, disulfide, amide and polyethylene glycol groups in any combination. In certain embodiments, each linking group is a linear aliphatic group comprising one or more groups selected from alkyl, substituted alkyl, ether, thioether, disulfide, amide, phosphodiester and polyethylene glycol groups in any combination. In certain embodiments, each linking group is a linear aliphatic group comprising one or more groups selected from alkyl, ether and amide groups in any combination. In certain embodiments, each linking group is a linear aliphatic group comprising one or more groups selected from alkyl, substituted alkyl, phosphodiester, ether and amide groups in any combination. In certain embodiments, each linking group is a linear aliphatic group comprising one or more groups selected from alkyl and phosphodiesters in any combination. In certain embodiments, each linking group comprises at least one phosphorus linking group or a neutral linking group.

In certain embodiments, the linking group comprises one or more cleavable bonds. In certain embodiments, the linking group is attached to the branching group through an amide or ether group. In certain embodiments, the linking group is attached to the branching group through a phosphodiester group. In certain embodiments, the linking group is attached to the branching group via a phosphorus linking group or a neutral linking group. In certain embodiments, the linking group is attached to the branching group through an ether group. In certain embodiments, the linking group is attached to the ligand through an amide or ether group. In certain embodiments, the linking group is attached to the ligand through an ether group. In certain embodiments, the linking group is attached to the ligand through an amide or ether group. In certain embodiments, the linking group is attached to the ligand through an ether group.

In certain embodiments, each linking group comprises a chain length of about 8 to about 20 atoms between the ligand and the branching group. In certain embodiments, each linking group comprises a chain length of from about 10 to about 18 atoms between the ligand and the branching group. In certain embodiments, each linking group comprises a chain length of about 13 atoms.

In certain embodiments, the linking group has a structure selected from:

wherein each n is, independently, 1 to 20; and

each p is 1 to about 6.

In certain embodiments, the linking group has a structure selected from:

.

.

In certain embodiments, the linking group has a structure selected from:

In the above formula, each n is independently 1 to 20.

In certain embodiments, the linking group has a structure selected from:

wherein L is a phosphorus linking group or a neutral linking group;

Z ₁ is C(=O)OR ₂ ;

Z ₂ is H, C ₁ -C ₆ alkyl or substituted C ₁ -C ₆ alkynyl (alky), and;

R ₂ is H, C ₁ -C ₆ alkyl or substituted C ₁ -C ₆ alkynyl (alky), and; and

each m ₁ is, independently, 0 to 20, wherein at least one m ₁ is greater than 0 for each linking group.

In certain embodiments, the linking group has a structure selected from:

.

.

In certain embodiments, the linking group has a structure selected from:

wherein Z ₂ is H or CH ₃ ; and

each m ₁ is, independently, 0 to 20, wherein at least one m ₁ is greater than 0 for each linking group.

In certain embodiments, the linking group has a structure selected from:

; 여기서 각각의 n은 독립적으로, 0, 1, 2, 3, 4, 5, 6, 또는 7이다.

; wherein each n is independently 0, 1, 2, 3, 4, 5, 6, or 7.

In certain embodiments, the linking group comprises a phosphorus linking group. In certain embodiments, the linking group does not include any amide linkages. In certain embodiments, the linking group includes a phosphorus linking group and does not include any amide linkages.

3. specific ligand

In certain embodiments, the present disclosure provides ligands, wherein each ligand is covalently bound to a linking group. In certain embodiments, each ligand that has affinity for at least one type of receptor on a target cell is selected. In certain embodiments, a ligand that has affinity for at least one type of receptor on the surface of a mammalian liver cell is selected. In certain embodiments, a ligand that has affinity for the hepatic asialoglycoprotein receptor (ASGP-R) is selected. In certain embodiments, each ligand is a carbohydrate. In certain embodiments, each ligand is independently selected from galactose, N-acetyl galactosamine, mannose, glucose, glucosamon, and fucose. In certain embodiments, each ligand is N-acetyl galactosamine (GalNAc). In certain embodiments, the targeting moiety comprises 2 to 6 ligands. In certain embodiments, the targeting moiety comprises three ligands. In certain embodiments, the targeting moiety comprises a 3 N-acetyl galactosamine ligand.

In certain embodiments, the ligand is a carbohydrate, carbohydrate derivative, modified carbohydrate, polyvalent carbohydrate cluster, polysaccharide, modified polysaccharide, or polysaccharide derivative. In certain embodiments, the ligand is an amino sugar or a thio sugar. For example, amino sugars can be used in any number of compounds known in the art, such as glucosamine, sialic acid, α-D-galactosamine, N-acetylgalactosamine, 2-acetamido-2-de oxy-D-galactopyranose (GalNAc), 2-amino-3- O -[( R )-1-carboxyethyl]-2-deoxy-β-D-glucopyranose (β-muramic acid), 2-deoxy-2-methylamino-L-glucopyranose, 4,6-dideoxy-4-formamido-2,3-di- O -methyl-D-mannopyranose, 2-deoxy -2-sulfoamino-D-glucopyranose and N -sulfo-D-glucosamine, and N -glycoloyl-α-neuraminic acid. For example, the thiosaccharide is 5-thio-β-D-glucopyranose, methyl 2,3,4-tri- O -acetyl-1-thio-6- O -trityl-α-D-glucopyrano side, 4-thio-β-D-galactopyranose, and ethyl 3,4,6,7-tetra- O -acetyl-2-deoxy-1,5-dithio-α-D- gluco -hep It may be selected from the group consisting of topiranoside.

In certain embodiments, “GalNac” or “Gal-NAc” refers to 2-(acetylamino)-2-deoxy-D-galactopyranose commonly referred to in the literature as N-acetyl galactosamine. In certain embodiments, “N-acetyl galactosamine” refers to 2-(acetylamino)-2-deoxy-D-galactopyranose. In certain embodiments, “GalNac” or “Gal-NAc” refers to 2-(acetylamino)-2-deoxy-D-galactopyranose. In certain embodiments, “GalNac” or “Gal-NAc” refers to β-type: 2-(acetylamino)-2-deoxy-β-D-galactopyranose and α-type: 2-(acetylamino) 2-(acetylamino)-2-deoxy-D-galactopyranose, including -2-deoxy-β-D-galactopyranose. In certain embodiments, β-type: 2-(acetylamino)-2-deoxy-β-D-galactopyranose and α-type: 2-(acetylamino)-2-deoxy-β-D- Both galactopyranose can be used interchangeably. Thus, in structures in which one form is depicted, these structures are intended to include the other form as well. For example, when a structure of α-type: 2-(acetylamino)-2-deoxy-D-galactopyranose is shown, this structure is intended to include other forms as well. In certain embodiments, in certain preferred embodiments, β-form 2-(acetylamino)-2-deoxy-D-galactopyranose is a preferred embodiment.

In certain embodiments the one or more ligands have a structure selected from:

wherein each R ₁ is selected from OH and NHCOOH.

In certain embodiments the one or more ligands have a structure selected from:

.

.

In certain embodiments the one or more ligands have a structure selected from:

.

.

In certain embodiments the one or more ligands have a structure selected from:

.

.

iii. specific conjugate

In certain embodiments, the conjugate group comprises the above structural features. In certain such embodiments, the conjugate group comprises the structure:

.

.

In the above formula, each n is independently 1 to 20.

In certain such embodiments, the conjugate group comprises the structure:

In certain such embodiments, the conjugate group has the structure:

In the above formula, each n is independently 1 to 20.

Z is H or a linked solid support;

Q is an antisense compound;

X is O or S; and

Bx is a heterocyclic base moiety.

In certain such embodiments, the conjugate group has the structure:

In certain such embodiments, the conjugate group has the structure:

In certain such embodiments, the conjugate group comprises the structure:

.

.

In certain such embodiments, the conjugate group comprises the structure:

.

.

In certain such embodiments, the conjugate group has the structure:

.

.

In certain such embodiments, the conjugate group has the structure:

.

.

In certain such embodiments, the conjugate group has the structure:

.

.

In certain such embodiments, the conjugate group has the structure:

.

.

In certain embodiments, the conjugate does not comprise pyrrolidine.

b. specific conjugated antisense compound

In certain embodiments, the conjugate is linked to the nucleoside of the antisense oligonucleotide at the 2', 3', or 5' position of the nucleoside. In certain embodiments, the conjugated antisense compound has the structure:

In the above formula,

A is an antisense oligonucleotide;

B is a cleavable moiety;

C is a conjugate linker

D is a branching group

each E is a linking group;

each F is a ligand; and

q is an integer from 1 to 5;

In certain embodiments, the conjugated antisense compound has the structure:

In the above formula,

A is an antisense oligonucleotide;

C is a conjugate linker

D is a branching group

each E is a linking group;

each F is a ligand; and

q is an integer from 1 to 5;

In certain such embodiments, the conjugate linker comprises at least one cleavable bond.

In certain such embodiments, the branching group comprises at least one cleavable bond.

In certain embodiments each linking group comprises at least one cleavable bond.

In certain embodiments, the conjugate is linked to the nucleoside of the antisense oligonucleotide at the 2', 3', or 5' position of the nucleoside.

In certain embodiments, the conjugated antisense compound has the structure:

In the above formula,

A is an antisense oligonucleotide;

B is a cleavable moiety;

C is a conjugate linker

each E is a linking group;

each F is a ligand; and

q is an integer from 1 to 5;

In certain embodiments, the conjugate is linked to the nucleoside of the antisense oligonucleotide at the 2', 3', or 5' position of the nucleoside. In certain embodiments, the conjugated antisense compound has the structure:

In the above formula,

A is an antisense oligonucleotide;

C is a conjugate linker

each E is a linking group;

each F is a ligand; and

q is an integer from 1 to 5;

In certain embodiments, the conjugated antisense compound has the structure:

In the above formula,

A is an antisense oligonucleotide;

B is a cleavable moiety;

D is a branching group

each E is a linking group;

each F is a ligand; and

q is an integer from 1 to 5;

In certain embodiments, the conjugated antisense compound has the structure:

In the above formula,

A is an antisense oligonucleotide;

D is a branching group

each E is a linking group;

each F is a ligand; and

q is an integer from 1 to 5;

In certain such embodiments, the conjugate linker comprises at least one cleavable bond.

In certain embodiments each linking group comprises at least one cleavable bond.

In certain embodiments, the conjugated antisense compound has a structure selected from:

.

.

In certain embodiments, the conjugated antisense compound has a structure selected from:

.

.

In certain embodiments, the conjugated antisense compound has a structure selected from:

.

.

In certain embodiments, the conjugated antisense compound has the structure:

.

.

Representative U.S. Patents, U.S. Patent Application Publications, and International Patents that teach the preparation of certain of the aforementioned conjugates, conjugated antisense compounds, linking groups, linkers, branching groups, ligands, cleavable moieties, as well as other modifications Application publications include, but are not limited to, US 5,994,517, US 6,300,319, US 6,660,720, US 6,906,182, US 7,262,177, US 7,491,805, US 8,106,022, US 7,723,509, US 2006/0148740, US 2011/0123520, WO 2013/033230 and WO 2012/037254, each of which is incorporated herein by reference in its entirety.

Representative publications teaching the preparation of certain of the aforementioned conjugates, conjugated antisense compounds, linking groups, linkers, branching groups, ligands, cleavable moieties, as well as other modifications include, but are not limited to: BIESSEN et al., "The Cholesterol Derivative of a Triantennary Galactoside with High Affinity for the Hepatic Asialoglycoprotein Receptor: a Potent Cholesterol Lowering Agent" J. Med. Chem. (1995) 38:1846-1852, BIESSEN et al., "Synthesis of Cluster Galactosides with High Affinity for the Hepatic Asialoglycoprotein Receptor" J. Med. Chem. (1995) 38:1538-1546, LEE et al., "New and more efficient multivalent glyco-ligands for asialoglycoprotein receptor of mammalian hepatocytes" Bioorganic & Medicinal Chemistry (2011) 19:2494-2500, RENSEN et al., "Determination of the Upper Size Limit for Uptake and Processing of Ligands by the Asialoglycoprotein Receptor on Hepatocytes in Vitro and in Vivo" J. Biol. Chem. (2001) 276(40):37577-37584, RENSEN et al., "Design and Synthesis of Novel N-Acetylgalactosamine-Terminated Glycolipids for Targeting of Lipoproteins to the Hepatic Asialoglycoprotein Receptor" J. Med. Chem. (2004) 47:5798-5808, SLIEDREGT et al., "Design and Synthesis of Novel Amphiphilic Dendritic Galactosides for Selective Targeting of Liposomes to the Hepatic Asialoglycoprotein Receptor" J. Med. Chem. (1999) 42:609-618, and Valentijn et al., “Solid-phase synthesis of lysine-based cluster galactosides with high affinity for the Asialoglycoprotein Receptor” Tetrahedron, 1997, 53(2), 759-770, (each of these) is incorporated herein by reference in its entirety).

In certain embodiments, the conjugated antisense compound is an RNase H based oligonucleotide (such as a gapmer) or a splice control oligonucleotide (such as a fully modified oligonucleotide) and any one comprising at least 1, 2, or 3 GalNAc groups. Contains conjugate groups. In certain embodiments the conjugated antisense compound comprises any of the conjugate groups found in any of the following references: Lee, Carbohydr Res , 1978, 67, 509-514; Connolly et al., J Biol Chem , 1982, 257, 939-945; Pavia et al., Int J Pep Protein Res , 1983, 22, 539-548; Lee et al., Biochem , 1984, 23, 4255-4261; Lee et al., Glycoconjugate J , 1987, 4, 317-328; Toyokuni et al., Tetrahedron Lett , 1990, 31, 2673-2676; Biessen et al., J Med Chem , 1995, 38, 1538-1546; Valentijn et al., Tetrahedron , 1997, 53, 759-770; Kim et al., Tetrahedron Lett , 1997, 38, 3487-3490; Lee et al., Bioconjug Chem , 1997, 8, 762-765; Kato et al., Glycobiol , 2001, 11, 821-829; Rensen et al., J Biol Chem , 2001, 276, 37577-37584; Lee et al., Methods Enzymol , 2003, 362, 38-43; Westerlind et al., Glycoconj J , 2004, 21, 227-241; Lee et al., Bioorg Med Chem Lett , 2006, 16(19), 5132-5135; Maierhofer et al., Bioorg Med Chem , 2007, 15, 7661-7676; Khorev et al., Bioorg Med Chem , 2008, 16, 5216-5231; Lee et al., Bioorg Med Chem , 2011, 19, 2494-2500; Kornilova et al., Analyt Biochem, 2012, 425, 43-46; Pujol et al., Angew Chemie Int Ed Engl , 2012, 51, 7445-7448; Biessen et al., J Med Chem , 1995, 38, 1846-1852; Sliedregt et al., J Med Chem , 1999, 42, 609-618; Rensen et al., J Med Chem , 2004, 47, 5798-5808; Rensen et al., Arterioscler Thromb Vasc Biol , 2006, 26, 169-175; van Rossenberg et al., Gene Ther , 2004, 11, 457-464; Sato et al., J Am Chem Soc , 2004, 126, 14013-14022; Lee et al., J Org Chem , 2012, 77, 7564-7571; Biessen et al., FASEB J , 2000, 14, 1784-1792; Rajur et al., Bioconjug Chem , 1997, 8, 935-940; Duff et al., Methods Enzymol , 2000, 313, 297-321; Maier et al., Bioconjug Chem , 2003, 14, 18-29; Jayaprakash et al., Org Lett , 2010, 12, 5410-5413; Manohran, Antisense Nucleic Acid Drug Dev , 2002, 12, 103-128; Merwin et al., Bioconjug Chem , 1994, 5, 612-620; Tomiya et al., Bioorg Med Chem, 2013, 21, 5275-5281; International application WO1998/013381; WO2011/038356; WO1997/046098; WO2008/098788; WO2004/101619; WO2012/037254; WO2011/120053; WO2011/100131; WO2011/163121; WO2012/177947; WO2013/033230; WO2013/075035; WO2012/083185; WO2012/083046; WO2009/082607; WO2009/134487; WO2010/144740; WO2010/148013; WO 1997/020563; WO2010/088537; WO2002/043771; WO2010/129709; WO2012/068187; WO2009/126933; WO2004/024757; WO2010/054406; WO2012/089352; WO2012/089602; WO2013/166121; WO2013/165816; US Patent 4,751,219; 8,552,163; 6,908,903; 7,262,177; 5,994,517; 6,300,319; 8,106,022; 7,491,805; 7,491,805; 7,582,744; 8,137,695; 6,383,812; 6,525,031; 6,660,720; 7,723,509; 8,541,548; 8,344,125; 8,313,772; 8,349,308; 8,450,467; 8,501,930; 8,158,601; 7,262,177; 6,906,182; 6,620,916; 8,435,491; 8,404,862; 7,851,615; Published US Patent Application Publications US2011/0097264; US2011/0097265; US2013/0004427; US2005/0164235; US2006/0148740; US2008/0281044; US2010/0240730; US2003/0119724; US2006/0183886; US2008/0206869; US2011/0269814; US2009/0286973; US2011/0207799; US2012/0136042; US2012/0165393; US2008/0281041; US2009/0203135; US2012/0035115; US2012/0095075; US2012/0101148; US2012/0128760; US2012/0157509; US2012/0230938; US2013/0109817; US2013/0121954; US2013/0178512; US2013/0236968; US2011/0123520; US2003/0077829; US2008/0108801; and US2009/0203132; (each of which is incorporated herein by reference in its entirety).

cell culture and antisense compound treatment

The effect of antisense compounds on the level, activity or expression of ANGPTL3 nucleic acids can be tested in vitro in a variety of cell types. Cell types used for such assays are available from commercial vendors (eg, American Type Culture Collection, Manassus, VA; Zen-Bio, Inc., Research Triangle Park, NC; Clonetics Corporation, Walkersville, MD). and cells are cultured according to the vendor's instructions using commercially available reagents (eg Invitrogen Life Technologies, Carlsbad, CA). Invitrogen Life Technologies, Carlsbad, CA). Exemplary cell types include, but are not limited to, HepG2 cells, Hep3B cells, Huh7 (hepatocellular carcinoma) cells, primary hepatocytes, A549 cells, GM04281 fibroblasts, and LLC-MK2 cells.

antisense of oligonucleotides in vitro exam

Described herein are methods of treatment of cells with antisense oligonucleotides that may be suitably modified for treatment with other antisense compounds.

Generally, cells are treated with antisense oligonucleotides when the cells reach approximately 60-80% confluency in culture.

One reagent commonly used to introduce antisense oligonucleotides into cultured cells includes the cationic lipid transfection reagent LIPOFECTIN® (Invitrogen, Carlsbad, CA). Antisense oligonucleotides were mixed with LIPOFECTIN® in OPTI-MEM® 1 (Invitrogen, Carlsbad, CA) to achieve the desired final concentration of antisense oligonucleotides and LIPOFECTIN® concentrations ranging from 2 to 12 ug/mL / 100 nM antisense oligonucleotides. do.

Another reagent used to introduce antisense oligonucleotides into cultured cells includes LIPOFECTAMINE 2000® (Invitrogen, Carlsbad, CA). Antisense oligonucleotides were prepared in LIPOFECTAMINE in OPTI-MEM® 1 reduced serum medium (Invitrogen, Carlsbad, CA) to achieve the desired concentration of antisense oligonucleotides and LIPOFECTAMINE® concentrations ranging from 2 to 12 ug/mL / 100 nM antisense oligonucleotides. mixed with 2000®.

Another reagent used to introduce antisense oligonucleotides into cultured cells includes Cytofectin® (Invitrogen, Carlsbad, CA). Antisense oligonucleotides were formulated with Cytofectin in OPTI-MEM® 1 reduced serum batch (Invitrogen, Carlsbad, CA) to achieve the desired concentration of antisense oligonucleotide and Cytofectin® concentrations ranging from 2 to 12 ug/mL / 100 nM antisense oligonucleotides. ® is mixed with

Another reagent used to introduce antisense oligonucleotides into cultured cells includes Oligofectamine™ (Life Technologies, Carlsbad, CA). Antisense oligonucleotides were prepared with Oligofectamine in Opti-MEM™-1 reduced serum medium (Life Technologies, Carlsbad, CA) to achieve the desired concentration of antisense oligonucleotides and Oligofectamine™ concentrations ranging from 0.2 to 0.8 μL / 100 nM antisense oligonucleotides. ™ is mixed with

Other reagents for introducing antisense oligonucleotides into cultured cells include FuGENE 6 (Roche Diagnostics Corp., Indianapolis, IN). Antisense oligomeric compounds were mixed with FuGENE 6 to 1-4 μL of oligomeric compounds FuGENE 6 (per 100 nM) FuGENE 6 in 1 mL serum-free RPMI to achieve the desired concentration of oligonucleotides.

Another technique used to introduce antisense oligonucleotides into cultured cells involves electroporation (Sambrook and Russell, Molecular Cloning: A Laboratory Manual, 3 ^rd Ed., 2001).

Cells are treated with antisense oligonucleotides by routine methods. Cells are typically harvested 16-24 hours after antisense oligonucleotide treatment, at which time RNA or protein levels of the target nucleic acid are measured by methods known in the art and described herein. In general, when treatments are performed in multiple replicates, data are provided as an average of replicate treatments.

The concentration of antisense oligonucleotide used varies from cell line to cell line. Methods for determining the optimal antisense oligonucleotide concentration for a particular cell line are known in the art. Antisense oligonucleotides are typically used at concentrations ranging from 1 nM to 300 nM when transfected with LIPOFECTAMINE2000®, Lipofectin or Cytofectin. Antisense oligonucleotides are used at high concentrations ranging from 625 to 20,000 nM when transfected using electroporation.

RNA isolation

RNA analysis can be performed on total cellular RNA or poly(A)+ mRNA. RNA isolation methods are well known in the art (Sambrook and Russell, Molecular Cloning: A Laboratory Manual, 3 ^rd Ed., 2001). RNA is prepared using methods known in the art, for example, TRIZOL® reagent (Invitrogen, Carlsbad, CA) according to the manufacturer's recommended protocol.

Inhibition assay of target level or expression

Inhibition of the level or expression of ANGPTL3 nucleic acid can be assayed in a variety of ways known in the art (Sambrook and Russell, Molecular Cloning: A Laboratory Manual, 3 ^rd Ed., 2001). For example, target nucleic acid levels can be quantified by, for example, Northern blot analysis, competitive polymerase chain reaction (PCR), or quantitative real-time PCR. RNA analysis can be performed on total cellular RNA or poly(A)+ mRNA. Methods of RNA isolation are known in the art. Northern blot analysis is also routine in the art. Quantitative real-time PCR can be conveniently accomplished using a commercially available ABI PRISM® 7600, 7700, or 7900 sequence detection system (available from PE-Applied Biosystems, Foster City, CA) and used according to the manufacturer's instructions. .

Quantitative real-time of target RNA levels PCR analyze

Quantification of target RNA levels can be achieved by quantitative real-time PCR using an ABI PRISM® 7600, 7700, or 7900 sequence detection system (PE-Applied Biosystems, Foster City, CA) according to the manufacturer's instructions. Methods of quantitative real-time PCR are known in the art.

Before real-time PCR, the isolated RNA is subjected to a reverse transcriptase (RT) reaction, which produces complementary DNA (cDNA) and is then used as a substrate for real-time PCR amplification. RT and real-time PCR reactions are performed sequentially in the same sample wells. RT and real-time PCR reagents are obtained from Invitrogen (Carlsbad, CA). RT, real-time-PCR reactions are performed by methods known to those skilled in the art.

Gene (or RNA) target amounts obtained by real-time PCR were calculated using expression levels of genes with constant expression, such as cyclophilin A or GADPH, or using RiboGreen® (Life Technologies ^™ , Inc. Carlsbad, Calif.) to total RNA can be quantified and normalized. Cyclophilin A or GADPH expression can be quantified by real-time PCR, concurrently with the target, multiplexed, or run separately. Total RNA can be quantified using RiboGreen® RNA Quantification Reagent. A method for RNA quantification by RiboGreen® is taught in Jones, LJ, et al, (Analytical Biochemistry, 1998, 265, 368-374). A CYTOFLUOR® 4000 instrument (PE Applied Biosystems) is used to measure RiboGreen® fluorescence.

Methods for designing real-time PCR probes and primers are known in the art and may include the use of software such as PRIMER EXPRESS® software (Applied Biosystems, Foster City, CA). Probes and primers used for real-time PCR are ANGPTL3-specific. designed to hybridize to sequences and are disclosed in the Examples below. A target specific PCR probe may have a FAM covalently linked to the 5' end and TAMRA or MGB covalently linked to the 3' end, wherein FAM is a fluorescent dye and TAMRA or MGB is a quencher dye .

Protein level analysis

Antisense inhibition of ANGPTL3 nucleic acids can be assessed by measuring ANGPTL3 protein levels. Protein levels of ANGPTL3 can be measured in a variety of ways known in the art, such as immunoprecipitation, Western blot analysis (immunoblotting), enzyme-linked immunosorbent assay (ELISA), quantitative protein assay, protein activity assay (e.g., caspase activity assays), immunohistochemistry, immunocytochemistry, or fluorescence-activated cell sorting (FACS) (Sambrook and Russell, Molecular Cloning: A Laboratory Manual, 3 ^rd Ed., 2001). Antibodies directed against a target can be identified and obtained from a variety of sources, such as the MSRS catalog of antibodies (Aerie Corporation, Birmingham, MI), or conventional methods for producing monoclonal or polyclonal antibodies well known in the art can be used. can be manufactured through

antisense compound in vivo exam

Antisense compounds, such as antisense oligonucleotides, are tested in animals to evaluate their ability to inhibit expression of ANGPTL3 and produce phenotypic changes. The test may be performed in normal animals or in experimental disease models. For administration to animals, antisense oligonucleotides are formulated in a pharmaceutically acceptable diluent such as phosphate-buffered saline. Administration includes parenteral routes of administration. Following a period of treatment with antisense oligonucleotides, RNA is isolated from the tissue and changes in ANGPTL3 nucleic acid expression are measured. Changes in ANGPTL3 protein levels are also measured.

specific signs

In certain embodiments, provided herein is a method of treating an individual comprising administering one or more pharmaceutical compositions as described herein. In certain embodiments, the subject has a metabolic disease and/or a cardiovascular disease. In certain embodiments, the subject has: hyperlipidemia (eg, familial or non-familial), hypercholesterolemia (eg, familial allogeneic hypercholesterolemia (HoFH), familial heterozygous hypercholesterolemia) (HeFH)), dyslipidemia, lipodystrophy, hypertriglyceridemia (e.g., heterologous LPL deficiency, allogeneic LPL deficiency), coronary artery disease (CAD), familial chylomicronemia syndrome (FCS), hyperlipidemia proteinemia type IV), metabolic syndrome, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), diabetes (eg type 2 diabetes, type 2 diabetes with dyslipidemia), insulin Resistance (eg, insulin resistance with dyslipidemia), vessel wall thickeners, hypertension (eg pulmonary arterial hypertension), sclerosis (eg, atherosclerosis, systemic sclerosis, progressive cutaneous sclerosis and proliferative obstructive angiopathy) eg digital ulcers and pulmonary vascular involvement) and combinations thereof.

In certain embodiments, compounds targeted to ANGPTL3 described herein modulate lipid and/or energy metabolism in animals. In certain embodiments, compounds targeted to ANGPTL3 described herein modulate physiological markers or phenotypes of hypercholesterolemia, dyslipidemia, lipodystrophy, hypertriglyceridemia, metabolic syndrome, NAFLD, NASH and/or diabetes. For example, administering the compound to an animal may modulate one or more of VLDL, non-esterified fatty acids (NEFA), LDL, cholesterol, triglycerides, glucose, insulin or ANGPTL3 levels. In certain embodiments, physiological markers or phenotypic modulation may be associated with inhibition of ANGPTL3 by the compound.

In certain embodiments, the compounds targeted to ANGPTL3 described herein reduce and/or prevent one or more of the following: hepatic TG accumulation (ie hepatic steatosis), atherosclerosis, thickening of blood vessel walls (eg, arterial Intima thickening), combined hyperlipidemia (eg, familial or non-familial), hypercholesterolemia (eg, familial allogeneic hypercholesterolemia (HoFH), familial heterozygous hypercholesterolemia (HeFH)) ), dyslipidemia, lipodystrophy, hypertriglyceridemia (e.g., heterologous LPL deficiency, allogeneic LPL deficiency, familial chylomicronemia syndrome (FCS), hyperlipoproteinemia type IV), metabolic syndrome, non- alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), diabetes (eg, type 2 diabetes, type 2 diabetes with dyslipidemia), insulin resistance (eg, with dyslipidemia) insulin resistance), hypertension and sclerosis, or any combination thereof. In certain embodiments, compounds targeted to ANGPTL3 described herein enhance insulin sensitivity.

In certain embodiments, administration of an antisense compound targeted to an ANGPTL3 nucleic acid by at least about 15%, by at least about 20%, by at least about 25%, by at least about 30%, by at least about 35%, by at least about 40% , by at least about 45%, by at least about 50%, by at least about 55%, by at least about 60%, by at least about 65%, by at least about 70%, by at least about 75%, by at least about 80%, at least by results in a decrease in ANGPTL3 expression by about 85%, by at least about 90%, by at least about 95%, or by at least about 99% or to a range defined by any two of these values.

In certain embodiments, a pharmaceutical composition comprising an antisense compound targeted to ANGPTL3 is used for the manufacture of a medicament for treating a patient suffering from or susceptible to a metabolic or cardiovascular disease. In certain embodiments, a pharmaceutical composition comprising an antisense compound targeting ANGPTL3 is used in the manufacture of a medicament for treating a patient suffering from, or susceptible to: combined hyperlipidemia (e.g., familial or non -familial), hypercholesterolemia (eg, familial allogeneic hypercholesterolemia (HoFH), familial heterozygous hypercholesterolemia (HeFH)), dyslipidemia, lipodystrophy, hypertriglyceridemia (eg, Familial chylomicronemia syndrome (FCS), hyperlipoproteinemia type IV), metabolic syndrome, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), diabetes (eg type 2) diabetes, type 2 diabetes with dyslipidemia), insulin resistance (eg, insulin resistance with dyslipidemia), vessel wall thickeners, hypertension and sclerosis, or any combination thereof.

administration

In certain embodiments, the compounds and compositions described herein are administered parenterally.

In certain embodiments, parenteral administration is by infusion. Infusions can be chronic, continuous, short-term or intermittent. In certain embodiments, the infused pharmaceutical agent is delivered by a pump.

In certain embodiments, parenteral administration is by injection. Such injections may be administered by syringe. In certain embodiments, parenteral administration is by injection. In certain embodiments, the injection is administered directly into a tissue or organ. In certain embodiments, the injection is administered subcutaneously.

any combination therapy

In certain embodiments, a first agent comprising a modified oligonucleotide is co-administered with one or more second agents. In certain embodiments, such second agents are designed to treat the same disease, disorder, or condition as the first agent described herein. In certain embodiments, such a second agent is designed to treat a different disease, disorder, or condition as the first agent described herein. In certain embodiments, such second agents are designed to treat undesirable side effects of one or more pharmaceutical compositions as described herein. In certain embodiments, the second agent is co-administered with the first agent to treat an unwanted effect of the first agent. In certain embodiments, the second agent is co-administered with the first agent to obtain a combination effect. In certain embodiments, the second agent is co-administered with the first agent to obtain a synergistic effect.

In certain embodiments, the first agent and one or more second agents are administered simultaneously. In certain embodiments, the first agent and the one or more second agents are administered at different times. In certain embodiments, a first agent and one or more second agents are prepared together in a single pharmaceutical formulation. In certain embodiments, the first agent and the one or more second agents are prepared separately.

In certain embodiments, the second agent includes, but is not limited to, a glucose-lowering agent. Glucose lowering agents include, but are not limited to, therapeutic lifestyle changes, PPAR agonists, dipeptidyl peptidase (IV) inhibitors, GLP-1 analogs, insulin or insulin analogs, insulin secretagogues, SGLT2 inhibitors, human amylin analogs, biguanides, an alpha-glucosidase inhibitor, or a combination thereof. Glucose-lowering agents may include, but are not limited to, metformin, sulfonylureas, rosiglitazone, meglitinide, thiazolidinediones, alpha-glucosidase inhibitors, or combinations thereof. The sulfonylurea may be acetohexamide, chlorpropamide, tolbutamide, tolazamide, glimepiride, glipizide, glyburide, or gliclazide. The meglitinide may be nateglinide or repaglinide. The thiazolidinedione may be pioglitazone or rosiglitazone. Alpha-glucosidase may be acarbose or miglitol. In certain embodiments, lipid lowering therapies may include, but are not limited to: therapeutic lifestyle changes, niacin, HMG-CoA reductase inhibitors, cholesterol absorption inhibitors, MTP inhibitors (eg, small molecules, polypeptides, antibodies or antisense compounds (targeted to MTP), fibrates, PCSK9 inhibitors (eg, PCSK9 antibodies, polypeptides, small molecule nucleic acid compounds (targeted to PCSK9)), CETP inhibitors (eg, small molecules such as torcetrapib and anacetrapib, polypeptide, antibody or nucleic acid compound (targeted to CETP), apoC-III inhibitor (e.g. small molecule, polypeptide, antibody or nucleic acid compound (targeted to apoC-III)), apoB inhibitor (e.g. For example, small molecules, polypeptides, antibodies or nucleic acid compounds (targeted to apoB), beneficial fats rich in omega-3 fatty acids, omega-3 fatty acids, or any combination thereof. The HMG-CoA reductase inhibitor may be atorvastatin, rosuvastatin, fluvastatin, lovastatin, pravastatin, simvastatin, or the like. The cholesterol absorption inhibitor may be ezetimibe. The fibrate may be fenofibrate, bezafibrate, ciprofibrate, clofibrate, gemfibrozil, or the like. Beneficial oils rich in omega-3 fatty acids may be krill, fish (eg Vascepa ^R ), flaxseed oil, and the like. The omega-3 fatty acid may be ALA, DHA, EPA, or the like.

specific compound

Antisense oligonucleotides targeting human ANGPTL3 have been described in previous publications (see PCT Patent Publication No. WO 2011/085271, published July 14, 2011, which is incorporated herein by reference in its entirety). Containing the strongest 10 antisense compounds best in vitro, an oligonucleotide can be one described in the above document nucleotides (233 676, 233 690, 233 710, 233 717, 233 721, 233 722, 337 459, 337 460, 337 474, 337 477, 337 478, 337 479, 337 481, 337484, 337487, 337488, 337490, 337491, 337492, 337497, 337498, 337503, 337505, 337506, 337508, 337513, 337514, 337516, 337520, 337521, 337525, 337526 and 337528) and U.S. Series Nos. 61/920,652 It was used as a benchmark throughout the selection in vitro screens for the described antisense compounds. Among the most potent compounds described in WO 2011/085271, ISIS 233722 was found to be highly variable in its ability to inhibit ANGPTL3. Therefore, 233722 was not selected as a benchmark for further studies, although initially included in some in vitro studies. Of the previously identified potent in vitro benchmark compounds, five (233710, 233717, 337477, 337478, 337479 and 337487) were selected for in vivo testing in huANGPTL3 transgenic mice, as described below, for human mRNA transcripts. and the most potent in reducing protein expression (Example 126). Antisense oligonucleotides with maximal initial in vivo efficacy in reducing ANGPTL3 levels (233710) were used as benchmarks for in vivo evaluation of novel antisense compounds described later.

In certain embodiments, the antisense compounds described herein benefit from one or more improved properties relative to the antisense compounds described in WO 2011/085271 and US Serial No. 61/920,652. These improved properties are demonstrated in the Examples herein, and a non-exhaustive summary of the Examples is provided below for ease of reference.

In the first screen described herein, about 3000 newly designed 5-10-5 MOE gapmer antisense compounds targeting human ANGPTL3 were tested in Hep3B cells for their effects on human ANGPTL3 mRNA in vitro ( Example 116). The level of mRNA inhibition of novel antisense compounds was assessed with several predesigned antisense compounds (233717, 337484, 337487, 337492 and 337516) used as benchmarks in selection studies. Out of about 3000 newly designed antisense compounds from this first screening, about 85 antisense compounds were selected for in vitro dose-dependent inhibition studies to determine 1/2 of their maximal inhibitory concentration (IC _{50 ) (} Examples 117-118). Of about 85 new antisense compounds tested for 1/2 of their maximal inhibitory concentration (IC ₅₀ ), about 38 antisense compounds demonstrating a strong dose-dependent reduction of ANGPTL3 were used as benchmarks, antisense compound 233710 was selected for in vivo efficacy and tolerability (ALT and AST) tests in mice (Examples 126-127).

In a second screen described herein, about 2000 newly designed antisense compounds targeting human ANGPTL3 with MOE gapmer motifs or mixed motifs (deoxy, 5-10-5 MOE and cET gapmers) were also tested in vitro with human Their effect on ANGPTL3 mRNA was tested in Hep3B cells (Examples 119-121). The level of inhibition of new antisense compounds was assessed with some previously designed antisense compounds (233717, 337487, 337513, 337514 and 337516) used as benchmarks in selection studies. Out of about 2000 newly designed antisense compounds from this second screening, about 147 antisense compounds were selected for in vitro dose-dependent inhibition studies to determine 1/2 of their maximal inhibitory concentration (IC _{50 ) (} Examples 122-125). Of about 147 new antisense compounds tested for 1/2 of their maximal inhibitory concentration (IC ₅₀ ), about 73 antisense compounds demonstrating a strong dose-dependent reduction in ANGPTL3 were used as benchmarks, antisense compound 233710 was selected for in vivo efficacy and tolerability (eg, ALT and AST) tests in mice (Examples 126-127).

Twenty-four of about 111 antisense compounds from screens 1 and 2 tested for efficacy and tolerability in mice were treated with alanine transaminase (ALT), aspartate transaminase (AST), albumin and bilirubin, and also renal metabolism. It was selected for more extensive tolerance testing in mice by evaluating markers BUN and liver metabolic markers such as creatinine and organ weight (Example 127).

In parallel with the in vivo murine study, 17 antisense compounds were selected for viscosity testing (Example 128). In general, antisense compounds that were not optimal for viscosity were not taken upwards in further studies.

Based on the results of the mouse tolerance study, 20 antisense compounds were selected for in vivo tolerance testing in rats (Example 129). In rats, the tolerability of compounds in vivo was determined by measuring liver metabolic markers such as ALT, AST, albumin and bilirubin, body weight and organ weight, and also renal metabolic markers such as BUN, creatinine and total protein/creatinine ratio.

Twenty antisense compounds tested in rats were also evaluated for cross-reactivity to the rhesus monkey ANGPTL3 gene sequence (Example 130). Although the antisense compound was tested in cynomolgus monkeys in this study, the cynomolgus monkey ANGPTL3 sequence was not available for comparison with the sequence of the full-length compound, so the sequence of the antisense compound was compared to that of the closely related rhesus monkey. compared. The sequences of eight antisense compounds were found to have 0-2 mismatches with the rhesus ANGPTL3 gene sequence and were further studied in cynomolgus monkeys (Example 130). Eight antisense compounds (ISIS 563580, ISIS 560400, ISIS 567320, ISIS 567321, ISIS 544199, ISIS 567233, ISIS 561011 and ISIS 559277) were tested for inhibition of ANGPTL3 mRNA and protein expression and also for resistance in monkeys. In resistance studies, body weight, hepatic metabolic markers (ALT, AST and bilirubin), renal metabolic markers (BUN and creatinine), hematological parameters (blood cell count, hemoglobin and hematocrit), and pro-inflammatory markers (CRP and C3) were evaluated. measured. In addition, by measuring the concentration of full-length oligonucleotides present in the liver and kidney, the ratio of full-length oligonucleotides in the kidney/liver was calculated.

The sequence of the potent and resistant antisense compound, ISIS 563580, measured in cynomolgus monkeys, as shown in Examples 113-115 and 131, was further modified with changes in the GalNAc conjugate and/or backbone chemistry, It was evaluated for increased potency.

Accordingly, provided herein are antisense compounds having one or more improved characteristics, for example, improved characteristics compared to the antisense compounds described in WO 2011/085271 and US Serial No. 61/920,652. In certain embodiments, provided herein are antisense compounds comprising a modified oligonucleotide as described herein that is targeted to, or specifically hybridized with, a region of any one of SEQ ID NOs: 1 and 2.

In certain embodiments, certain antisense compounds as described herein are effective due to their efficacy in inhibiting ANGPTL3 expression. In certain embodiments, the compound or composition comprises at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85% ANGPTL3. , inhibit by at least 90% or at least 95%.

In certain embodiments, certain antisense compounds described herein are less than 20 μM, less than 10 μM, 8 μM when tested in human cells, e.g., the Hep3B cell line (as described in Examples 117-118 and 122-125). It is effective due to an in vitro IC ₅₀ of less than, less than 5 μM, less than 2 μM, less than 1 μM, less than 0.9 μM, less than 0.8 μM, less than 0.7 μM, less than 0.6 μM, or less than 0.5 μM. In certain embodiments, _{preferred antisense compounds having an IC 50} of less than 1.0 μM comprise SEQ ID NOs: 15, 20, 24, 34, 35, 36, 37, 42, 43, 44, 47, 50, 51, 57, 58, 60, 77, 79, 82, 87, 88, 90, 91, 93, 94, 100, 101, 104, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 169, 170, 177, 188, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, and 232. In certain embodiments, preferred antisense compounds having _{an IC 50 of less than 0.9 μM are} 15, 20, 35, 36, 42, 43, 44, 50, 57, 60, 77, 79, 87, 88, 90, 91, 93, 94, 101, 104, 110, 111 , 112, 113, 114, 115, 116, 117, 118, 119, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138 , 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 177, 209, 210, 211, 212 , 213, 214, 215, 216, 217, 218, 219, 22 0, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, and 232. In certain embodiments, _{preferred antisense compounds having an IC 50} of less than 0.8 μM include SEQ ID NOs: : 15, 20, 35, 36, 42, 43, 44, 50, 57, 60, 77, 79, 87, 88, 90, 91, 93, 94, 101, 104, 110, 111, 112, 113, 114 , 115, 116, 117, 118, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142 , 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 177, 209, 210, 211, 212, 213, 214, 215, 217 , 218, 219, 220, 221, 222, 223, 224, 225, 228, 229, 230, 231, and 232. In certain embodiments, _{preferred antisense compounds having an IC 50} of less than 0.7 μM include SEQ ID NOs: Do: 15, 20, 36, 42, 43, 57, 60, 114, 117, 127, 131, 177, 209, 210, 211, 212, 213, 214, 215, 217, 218, 219, 220, 221, 222, 223, 224, 225, 228, 229, 230, 231, and 232. In certain embodiments, _{preferred antisense compounds having an IC 50} of less than 0.6 μM include SEQ ID NOs: 15, 20, 36, 42 , 43, 57, 60, 114, 117, 127, 131, 177, 209, 210, 211, 212, 213, 215, 217, 218, 219, 220, 221, 222, 224, 225, 228, 229, 230 , 231, and 232. In certain embodiments, 0.5 μ _{Preferred antisense compounds having an IC 50} of less than M include SEQ ID NOs: 43, 114, 117, 127, 131, 177, 209, 210, 211, 212, 215, 217, 218, 219, 220, 221, 222, 229, 230, and 232.

In certain embodiments, certain antisense compounds as described herein have a viscosity of less than 40 cP, less than 35 cP, less than 30 cP, less than 25 cP, as measured by an assay (as described in Example 128); It is effective because it is less than 20 cP, less than 15 cP, or less than 10 cP. Oligonucleotides with a viscosity greater than 40 cP will have a less than optimal viscosity. In certain embodiments, preferred antisense compounds having a viscosity of less than 20 cP comprise the following SEQ ID NOs: 16, 18, 20, 34, 35, 36, 38, 49, 77, 90, 93, and 94. Certain embodiments In an example, preferred antisense compounds having a viscosity of less than 15 cP include the following SEQ ID NOs: 16, 18, 20, 34, 35, 38, 49, 90, 93, and 94. In certain embodiments, less than 10 cP Preferred antisense compounds having a viscosity of include SEQ ID NOs: 18, 34, 35, 49, 90, 93, and 94.

In certain embodiments, certain antisense compounds as described herein are highly resistant, as evidenced by the in vivo tolerance measurements described in the Examples. In certain embodiments, certain antisense compounds as described herein have an increase in ALT and/or AST values of no more than 3-fold, 2-fold, or 1.5-fold over saline treated animals, as evidenced by: It is highly resistant.

In certain embodiments, certain antisense compounds as described herein have an inhibitory potency of at least 50%, an in vitro IC ₅₀ of less than 1 μM, a viscosity of less than 20 cP, and a 3-fold or less increase in ALT and/or AST. It is effective due to having more than one.

In certain embodiments, ISIS 563580 (SEQ ID NO: 77) is preferred. This compound was found to be a potent inhibitor and the most resistant antisense compound in ANGPTL3 transgenic mice. It had an acceptable viscosity of about 16.83 cP and _{an IC 50} value of less than 0.8 μM in vitro. In mice, it was up to 3-fold increased in ALT and/or AST levels than in saline treated animals. Also, in monkeys, it was among the most resistant and potent compounds in inhibiting ANGPTL3 and had the best ratio of full-length oligonucleotide concentrations.

In certain embodiments, ISIS 544199 (SEQ ID NO: 20) is preferred. It has been found that this compound is a potent and tolerable antisense compound. It had an acceptable viscosity of about 1.7 cP and _{an IC 50} value of less than 0.5 μM in vitro. In mice, it was up to 3-fold increased in ALT and/or AST levels than in saline treated animals. Also, in monkeys, it was among the most potent compounds in inhibiting ANGPTL3 and had a good ratio of full-length oligonucleotide concentrations.

In certain embodiments, ISIS 559277 (SEQ ID NO: 110) is preferred. It has been found that this compound is a potent and tolerable antisense compound. This resulted in an IC ₅₀ value of less than 0.8 μM in vitro. In mice, it was up to 3-fold increased in ALT and/or AST levels than in saline treated animals. Also, in monkeys, it was among the most potent compounds in inhibiting ANGPTL3 and had a good ratio of full-length oligonucleotide concentrations.

In certain embodiments, a GalNAc conjugated antisense compound, ISIS 658501 (SEQ ID NO: 4912) is preferred. This antisense compound was shown to be more potent (as indicated by the level of inhibition) than its parent compound ISIS 563580 (SEQ ID NO: 77).

In certain embodiments, a GalNAc conjugated antisense compound, ISIS 703801 (SEQ ID NO: 77) is preferred. This antisense compound was shown to be more potent than its parent compound ISIS 563580 (SEQ ID NO: 77). ISIS 703801 has an ID50 value of 1, while ISIS 563580 has an ID50 value of 6.

In certain embodiments, a GalNAc conjugated antisense compound, ISIS 703802 (SEQ ID NO: 77) is preferred. This antisense compound was shown to be more potent than its parent compound ISIS 563580 (SEQ ID NO: 77). ISIS 703802 has an ID50 value of 0.3, while ISIS 563580 has an ID50 value of 6.

Example

The following examples illustrate and do not limit specific embodiments of the present disclosure. Moreover, where specific embodiments are provided, the inventors have contemplated generic applications of the specific embodiments. For example, the disclosure of an oligonucleotide having a particular motif provides reasonable support for additional oligonucleotides having the same or similar motif. And, for example, if a particular high-affinity modification occurs at a particular position, other high-affinity modifications at the same position are considered suitable unless otherwise indicated.

Example One: phosphoramidite , General Methods for the Preparation of Compounds 1, 1a, and 2

Compounds 1, 1a and 2 were prepared according to procedures well known in the art, as described herein (Seth et al., Bioorg. Med. Chem., 2011, 21(4), 1122-1125). , J. Org. Chem., 2010, 75(5), 1569-1581, Nucleic Acids Symposium Series, 2008, 52(1), 553-554); and See also Published PCT International Applications (WO 2011/115818, WO 2010/077578, WO2010/036698, WO2009/143369, WO 2009/006478, and WO 2007/090071), and US Patent 7,569,686).

Example 2: Preparation of compound 7

Compound 3 (2-acetamido-1,3,4,6-tetra-O-acetyl-2-deoxy-β-Dgalactopyranose or galactosamine pentaacetate) is commercially available. Compound 5 was prepared according to published procedures (Weber et al., J. Med. Chem ., 1991, 34 , 2692).

Example 3: Preparation of compound 11

Compounds 8 and 9 are commercially available.

Example 4: Preparation of compound 18

Compound 11 was prepared according to the procedure demonstrated in Example 3. Compound 14 is commercially available. Compound 17 was prepared using a similar procedure reported by Rensen et al., J. Med . Chem ., 2004, 47, 5798-5808.

Example 5: Preparation of compound 23

Compounds 19 and 21 are commercially available.

Example 6: Preparation of compound 24

Compounds 18 and 23 were prepared according to the procedures demonstrated in Examples 4 and 5.

Example 7: compound 25 manufacturing

Compound 24 was prepared according to the procedure demonstrated in Example 6.

Example 8: Preparation of compound 26

Compound 24 is prepared according to the procedure demonstrated in Example 6.

Example 9: GalNAc ₃ - 1 containing at the 3' end conjugated ASO's General preparation, compound 29

The protected GalNAc ₃ - 1 has the following structure:

Konju gate group GalNAc ₃ -1 GalNAc ₃ Cluster moieties (GalNAc ₃ −1 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. wherein GalNAc ₃ -1 _a has the formula:

Solid support bound protected GalNAc ₃ -1 , compound 25 was prepared according to the procedure demonstrated in Example 7. Oligomeric compound 29 containing GalNAc ₃ -1 at the 3' end was prepared using standard procedures in automated DNA/RNA synthesis (Dupouy et al., Angew . Chem . Int . Ed., 2006, 45, 3623-3627). The phosphoramidite building blocks, compounds 1 and 1a, were prepared according to the procedure demonstrated in Example 1. The phosphoramidite demonstrated is representative and is not intended to be limiting as other phosphoramidite building blocks can be used to prepare oligomeric compounds of the desired order and composition. The order and amount of phosphoramidite added to the solid support can be adjusted to prepare the gapped oligomeric compounds described herein. Such gapped oligomeric compounds can have the desired composition and base sequence as dictated by any given target.

Example 10: GalNAc ₃ - 1 containing at the 5' end conjugated ASO's General preparation, compound 34

Unylinker ^TM 30 is commercially available. Oligomeric compound 34 containing GalNAc ₃ -1 at the 5' end was prepared using standard procedures in automated DNA/RNA synthesis (Dupouy et al., Angew . Chem . Int . Ed., 2006, 45, 3623-3627). The phosphoramidite building blocks, compounds 1 and 1a, were prepared according to the procedure demonstrated in Example 1. The phosphoramidite demonstrated is representative and is not intended to be limiting as other phosphoramidite building blocks can be used to prepare oligomeric compounds of the desired order and composition. The order and amount of phosphoramidite added to the solid support can be adjusted to prepare the gapped oligomeric compounds described herein. Such gapped oligomeric compounds can have the desired composition and base sequence as dictated by any given target.

Example 11: compound 39 manufacturing

Compounds 4, 13 and 23 were prepared according to the procedures demonstrated in Examples 2, 4, and 5. Compound 35 is prepared using a similar procedure known from Rouchaud et al., Eur. J. Org. Chem., 2011, 12 , 2346-2353.

Example 12: compound 40 manufacturing

Compound 38 is prepared according to the procedure demonstrated in Example 11.

Example 13: compound 44 manufacturing

Compounds 23 and 36 are prepared according to the procedures demonstrated in Examples 5 and 11. Compound 41 is prepared using a similar procedure known in WO 2009082607.

Example 14: compound 45 manufacturing

Compound 43 is prepared according to the procedure demonstrated in Example 13.

Example 15: compound 47 manufacturing

Compound 46 is commercially available.

Example 16: compound 53 manufacturing

Compounds 48 and 49 are commercially available. Compounds 17 and 47 are prepared according to the procedures demonstrated in Examples 4 and 15.

Example 17: compound 54 manufacturing

Compound 53 is prepared according to the procedure demonstrated in Example 16.

Example 18: compound 55 manufacturing

Compound 53 is prepared according to the procedure demonstrated in Example 16.

Example 19: 3 ' In position through solid-state technology GalNAc ₃ containing -1 conjugated General Method of Preparation of ASO (Production of ISIS 647535, 647536 and 651900)

Unless otherwise stated, all reagents and solutions used for the synthesis of oligomeric compounds are purchased from commercial sources. Standard phosphoramidite building blocks and solid supports are used for incorporation of nucleoside residues including, for example, T, A, G, and ^{m C residues.} A 0.1 M solution of phosphoramidite in anhydrous acetonitrile was used for β-D-2'-deoxyribonucleoside and 2'-MOE.

올리고파일럿 합성기 (40-200 μmol 규모) 상에서 수행되었다. 커플링 단계의 경우, 포스포르아미다이트를 고체 지지체의 로딩에 대해 4배 과량으로 전달하였고, 포스포르아미다이트 응축을 10분 동안 수행하였다. 모든 다른 단계는 제조자에 의해 공급된 표준 프로토콜을 따랐다. 톨루엔 중 6% 디클로로아세트산의 용액은 디메톡시트리틸 (DMT) 그룹을 뉴클레오티드의 5'-하이드록실 그룹로부터 제거하기 위해 사용되었다. 무수 CH₃CN 중 4,5-디시아노이미다졸 (0.7 M)은 커플링 단계 동안에 활성제로서 사용되었다. 포스포로티오에이트 연결을 3분의 접촉 시간 동안에 1:1 피리딘/CH3CN 중의 크산탄 하이드라이드의 0.1 M 용액을 이용한 황화에 의해 도입하였다. 6% 물을 함유하는 CH₃CN 중의 20% tert-부틸하이드로퍼옥사이드의 용액을 산화제로서 사용하여 12분의 접촉 시간으로 포스포디에스테르 뉴클레오시드간 연결을 제공하였다. ASO synthesis was performed in a phosphoramidite coupling method on an ABI 394 synthesizer (1-2 μmol scale) or on a column-packed GalNAc ₃ -1 loaded VIMAD solid support (110 μmol/g, Guzaev et al., 2003). by GE Healthcare Bioscience

performed on an oligopilot synthesizer (40-200 μmol scale). For the coupling step, phosphoramidite was delivered in a 4-fold excess relative to the loading of the solid support, and phosphoramidite condensation was carried out for 10 minutes. All other steps followed the standard protocol supplied by the manufacturer. A solution of 6% dichloroacetic acid in toluene was used to remove the dimethoxytrityl (DMT) group from the 5'-hydroxyl group of the nucleotide. 4,5-dicyanoimidazole (0.7 M) in anhydrous CH ₃ CN was used as activator during the coupling step. Phosphorothioate linkages were introduced by sulfiding with a 0.1 M solution of xanthan hydride in 1:1 pyridine/CHCN for a contact time of 3 minutes. A solution of 20% tert _{-butylhydroperoxide in CH 3} CN containing 6% water was used as the oxidizing agent to provide phosphodiester internucleoside linkages with a contact time of 12 minutes.

After the desired sequence was assembled, the cyanoethyl phosphate protecting group was deprotected using a 1:1 (v/v) mixture of triethylamine and acetonitrile with a contact time of 45 min. The solid-support bound ASO was suspended in aqueous ammonia (28-30 wt %) and heated at 55° C. for 6 hours.

Then, the unbound ASO was filtered off and the ammonia was removed by heating. The residue was purified by high pressure liquid chromatography on a strong anion exchange column (GE Healthcare Bioscience, Source 30Q, 30 μm, 2.54 x 8 cm, A = _{100 mM ammonium acetate in 30% aqueous CH 3} CN, B = in A 1.5 M NaBr, 0-40% B at 60 min, flow rate 14 mL min-1, λ = 260 nm). The residue was desalted by HPLC on a reversed phase column to give the desired ASO in isolated yields of 15-30% based on the initial loading on the solid support. ASO was characterized by ion-pair-HPLC coupled MS analysis with an Agilent 1100 MSD system.

Antisense oligonucleotides without conjugates were synthesized using standard oligonucleotide synthesis procedures well known in the art.

Using these methods, three distinct antisense compounds targeting ApoC III were prepared. As summarized in Table 17 below, the three antisense compounds targeting each ApoC III had the same nucleobase sequence; ISIS 304801 is a 5-10-5 MOE gapmer with all phosphorothioate linkages; ISIS 647535 is identical to ISIS 304801 except that it has GalNAc ₃ -1 conjugated at the 3'end; and ISIS 647536 is the same as ISIS 647535, provided that the specific internucleoside linkage of the compound is a phosphodiester linkage. As further summarized in Table 17, two separate antisense compounds targeting SRB-1 were synthesized. ISIS 440762 was a 2-10-2 cEt gapmer with all phosphorothioate internucleoside linkages; ISIS 651900 was identical to ISIS 440762, except that GalNAc ₃ -1 was included at the 3'-end.

Table 17

Modified ASOs targeting ApoC III and SRB-1

Subscript: Subscript: "e" represents a 2'-MOE modified nucleoside; "d" represents β-D-2'-deoxyribonucleoside;"k" represents a 6'- (S) -CH ₃ bicyclic nucleoside (eg cEt); "s" represents a phosphorothioate internucleoside linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO); And "o'" represents -OP(=O)(OH)-. The superscript "m" represents 5-methylcytosine. “ GalNAc ₃ -1 ” refers to a group of conjugates having the structure shown previously in Example 9. Note that GalNAc ₃ -1 contains a cleavable adenosine linking the ASO to the remainder of the conjugate designated as “GalNAc ₃ -1 _a ”. This nomenclature refers to the complete nucleobase sequence including atenosine that is part of the conjugate. is used in the table above to show. Thus, in the table above, sequences could also be listed as ending with " A _do " omitted with "GalNAc ₃ - 1". Cleavable nucleosides or cleavage This convention of using the subscript "a" to denote a portion of a conjugate group without a cleavable moiety is used throughout these examples.This portion of a conjugate group without a cleavable moiety is referred to as a "cluster" or referred to herein as a “conjugate cluster” or “GalNAc ₃ cluster.” In certain instances it is convenient to describe a conjugate group by separately providing its cluster and its cleavable moiety.

Example 20: huApoC III Transgenic Mice to Humans ApoC III Dose-dependent antisense inhibition

ISIS 304801 and ISIS 647535, which each target human ApoC III and described above, were tested separately and evaluated in a dose-dependent study for their ability to inhibit human ApoC III in human ApoC III transgenic mice.

cure

Human ApoCIII transgenic mice were maintained on a 12-hour light/dark cycle and were ad libitum fed to a Teklad laboratory cage. Animals were acclimatized for at least 7 days in the study facility prior to initiation of the experiment. ASO was prepared in PBS and sterilized by filtration through a 0.2 micron filter. ASO was dissolved in 0.9% PBS for injection.

Human ApoC III transgenic mice were injected intraperitoneally with ISIS 304801 or 647535 at 0.08 and 0.25 once a week for 2 weeks. 0.75, 2.25 or 6.75 μmol/kg or PBS as control. Each treatment group consisted of 4 animals. Forty-eight hours after administration of the last dose, blood was drawn from each mouse and the mice were sacrificed and tissues collected.

ApoC III mRNA analyze

ApoC III mRNA levels in the liver of mice were measured using real-time PCR and RIBOGREEN® RNA quantification reagent according to standard protocols (Molecular Probes, Inc. Eugene, OR). ApoC III mRNA levels were determined for total RNA (using ribogrin) prior to normalization to PBS-treated controls. Results below are presented as mean percent rover of ApoC III mRNA levels for each treatment group, normalized to PBS-treated controls, and are expressed as “% PBS”. The half maximal effective dose (ED ₅₀ ) of each ASO is also provided in Table 18 below.

As demonstrated, both antisense compounds reduced ApoC III RNA relative to the PBS control. In addition, GalNAc ₃ -1 conjugate the antisense compound to (ISIS 647535) is GalNAc ₃ -1 conjugate is substantially stronger than the antisense compounds do not have a (ISIS 304801).

Table 18

Effect of ASO Treatment on ApoC III mRNA Levels in Human ApoC III Transgenic Mice

ApoC III Protein Analysis ( turbidity ( Turbidometric ) black)

Plasma ApoC III protein analysis was determined using the procedure reported by Graham et al., Circulation Research, published online prior to print, March 29, 2013.

Approximately 100 μl of plasma isolated from mice was analyzed without dilution using an Olympus clinical analyzer and a commercially available nephrotic ApoC III assay (Kamiya, Cat# KAI-006, Kamiya Biomedical, Seattle, WA). The assay protocol was performed as described by the vendor.

As shown in Table 19 below, both antisense compounds reduced the ApoC III protein relative to the PBS control. In addition, GalNAc ₃ -1 conjugate the antisense compound to (ISIS 647535) is GalNAc ₃ -1 conjugate is substantially stronger than the antisense compounds do not have a (ISIS 304801).

Table 19

Effect of ASO Treatment on ApoC III Plasma Protein Levels in Human ApoC III Transgenic Mice

Plasma triglycerides and cholesterol are Bligh, EG and Dyer, WJ Can. J. Biochem. Physiol . 37: 911-917, 1959) (Bligh, E and Dyer, W, Can J Biochem). Physiol , 37, 911-917, 1959) (Bligh, E and Dyer, W, Can J Biochem Physiol , 37, 911-917, 1959), using a Beckmann Coulter clinical analyzer and commercially available reagents. was measured.

Triglyceride levels were measured for PBS-injected mice and are expressed as “% PBS”. Results are provided in Table 20. As demonstrated, both antisense compounds reduced triglyceride levels. In addition, GalNAc ₃ -1 conjugate the antisense compound to (ISIS 647535) is GalNAc ₃ -1 conjugate is substantially stronger than the antisense compounds do not have a (ISIS 304801).

Table 20

Effect of ASO Treatment on Triglyceride Levels in Transgenic Mice

Plasma samples were analyzed by HPLC to determine the amounts of total cholesterol and different fractions of cholesterol (HDL and LDL). Results are provided in Tables 21 and 22. As demonstrated, both antisense compounds reduced total cholesterol levels; Both reduced LDL; And both raised HDL. In addition, GalNAc ₃ -1 conjugate the antisense compound to (ISIS 647535) is GalNAc ₃ -1 conjugate is substantially stronger than the antisense compounds do not have a (ISIS 304801). An increase in HDL and a decrease in LDL levels are cardiovascular beneficial effects of antisense inhibition of ApoC III.

Table 21

Effect of ASO Treatment on Total Cholesterol Levels in Transgenic Mice

Table 22

Effect of ASO Treatment on HDL and LDL Cholesterol Levels in Transgenic Mice

Pharmacokinetic analysis (PK)

The PK of ASO was also assessed. Liver and kidney samples were minced and extracted using standard protocols. Samples were analyzed on MSD1 using IP-HPLC-MS. Tissue levels (μg/g) of full-length ISIS 304801 and 647535 were measured and the results are provided in Table 23. As demonstrated, liver concentrations of total full-length antisense compounds were similar for the two antisense compounds. Thus, although the GalNAc ₃ -1 conjugated antisense compound group is more active in the liver (as substantiated by the RNA and protein data above), it is not present in substantially higher concentrations in the liver. In fact, the calculated EC ₅₀ (provided in Table 23) confirms that the observed increase in potency of the conjugated compounds cannot be entirely attributable to the increased accumulation. These results suggest that the conjugates possibly improved the productive uptake of antisense compounds into cells, thereby improving efficacy by mechanisms other than hepatic accumulation alone.

The results also show that the concentration of the GalNAc ₃ -1 conjugated antisense compound in the kidney is lower than that of the antisense compound without the GalNAc conjugate. It has several beneficial therapeutic implications. For therapeutic indications where activity in the kidney is not sought, exposure to the kidney jeopardizes renal toxicity without a corresponding benefit. In addition, high concentrations in the kidneys typically cause a loss of the compound into the urine, thereby gaining faster clearance. Thus, for non-renal targets, renal accumulation is undesirable. These data suggest that GalNAc ₃ -1 conjugation reduces renal accumulation.

Table 23

PK analysis of ASO treatment in transgenic mice

A metabolite of ISIS 647535 was also identified, and its mass was confirmed by high-resolution mass spectrometry analysis. The cleavage sites and structures of the observed metabolites are shown below. The relative % of full-length ASO was calculated using standard procedures and the results are provided in Table 23a. The main subject of ISIS 647535 was a full-length ASO without the entire conjugate (ie ISIS 304801) that resulted in cleavage at cleavage site A shown below. In addition, additional objects obtained from other cleavage sites were also observed. These results indicate that other cleavable bonds such as esters, peptides, disulfides, phosphoramidates or acyl-hydrazones can be combined with GalNAc ₃ - 1 sugars and ASOs, which can be cleaved by enzymes in the cell or in the reducing environment of the cytosol. It also suggests that introduction between endosomes and lysosomes (labile to acidic pH) may also be useful.

Table 23a

Observed full-length metabolites of ISIS 647535

Example 21: Humans in a single dose study ApoC III Transgenic Mice to Humans ApoC III's antisense control

ISIS 304801, 647535 and 647536, each of which target human ApoC III and are described in Table 17, were further evaluated in a single dose study for their ability to inhibit human ApoC III in human ApoC III transgenic mice.

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Human ApoCIII transgenic mice were maintained on a 12-hour light/dark cycle and were ad libitum fed to a Teklad laboratory cage. Animals were acclimatized for at least 7 days in the study facility prior to initiation of the experiment. ASO was prepared in PBS and sterilized by filtration through a 0.2 micron filter. ASO was dissolved in 0.9% PBS for injection.

Human ApoC III transgenic mice were injected intraperitoneally once with ISIS 304801, 647535 or 647536 (described above) or PBS treated controls at the doses shown below. The treatment group consisted of 3 animals and the control group consisted of 4 animals. Before treatment as well as after the last dose, blood was drawn from each mouse and plasma samples were analyzed. Mice were sacrificed 72 hours after the last dose.

A sample was collected and analyzed to include liver containing HDL and LDL fractions; plasma triglycerides; and ApoC III mRNA and protein levels in cholesterol were measured and assessed as described above (Example 20). Data from these analyzes are provided in Tables 24-28 below. Serum hepatic transaminase levels, alanine transaminase (ALT) and aspartate transaminase (AST) were measured for saline infused mice using standard protocols. ALT and AST levels showed that the antisense compound was tolerated at all administered doses.

These results show an improvement in efficacy for antisense compounds containing GalNAc ₃ -1 conjugates at the 3' end (ISIS 647535 and 647536) compared to antisense compounds without GalNAc ₃ -1 conjugates (ISIS 304801). Moreover, ISIS 647536 with the GalNAc ₃ -1 conjugate and some phosphodiester linkages was as potent as ISIS 647535 with the same conjugate, and all internucleoside linkages in ASO are phosphorothioate.

Table 24

Effect of ASO Treatment on ApoC III mRNA Levels in Human ApoC III Transgenic Mice

Table 25

Effect of ASO Treatment on ApoC III Plasma Protein Levels in Human ApoC III Transgenic Mice

Table 26

Effect of ASO Treatment on Triglyceride Levels in Transgenic Mice

Table 27

Effect of ASO Treatment on Total Cholesterol Levels in Transgenic Mice

Table 28

Effect of ASO Treatment on HDL and LDL Cholesterol Levels in Transgenic Mice

These results confirm that the GalNAc ₃ -1 conjugate improves the efficacy of antisense compounds. The results also show equivalent efficacy of GalNAc ₃ -1 conjugated antisense compounds, wherein the antisense oligonucleotides have mixed linkages (ISIS 647536 with 6 phosphodiester linkages) and complete phosphorothioate of the same antisense compound. version (ISIS 647535).

Phosphorothioate linkages provide several properties for antisense compounds. For example, in nuclease digestion, it stores and binds to proteins, which causes accumulation of the compound in the liver rather than in the kidneys/urine. These are desirable properties, especially when treating aura in the liver. However, phosphorothioate linkages have also been associated with inflammatory responses. Thus, reducing the number of phosphorothioate linkages in a compound reduces the risk of inflammation, as well as low concentrations of the compound in the liver, increases its concentrations in the kidneys and urine, increases its stability in the presence of nucleases and , is expected to lower the overall efficacy lower. These results show that the GalNAc ₃ -1 conjugated antisense compound in which a specific phosphorothioate linkage has been replaced by a phosphodiester linkage is potent against the target in the liver as a counterpart with a complete phosphorothioate linkage. Such compounds are described as being less pro-inflammatory (see Example 24, which describes that experiments showing a reduction in PS resulted in reduced inflammatory effects).

Example 22: in vivo at SRB targeting -1 GalNAc ₃ -One conjugated deformed ASO's effect

Its ability to target SRB-1 and inhibit SRB-1 in Balb/c mice was evaluated in a dose-dependent study for each of ISIS 440762 and 651900 described in Table 17.

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6-week-old male Balb/c mice (Jackson Laboratory, Bar Harbor, ME) were injected subcutaneously once with ISIS 440762, 651900 or PBS-treated controls at the doses given below. Each treatment group consisted of 4 animals. Mice were sacrificed 48 h after the final dose and SRB-1 mRNA levels in the liver were measured using real-time PCR and RiboGreen® RNA quantitation reagent according to standard protocols (Molecular Probes, Inc. Eugene, OR). SRB-1 mRNA levels were determined for total RNA (using ribogreen) prior to normalization to PBS-treated controls. Results below are presented as average percent of SRB-1 mRNA levels for each treatment group, normalized to PBS-treated controls, and are expressed as “% PBS”.

As demonstrated in Table 29, both antisense compounds lowered SRB-1 mRNA levels. Moreover, the antisense compound containing the GalNAc ₃ -1 (ISIS 651900) conjugate was substantially stronger than the antisense compound without the GalNAc ₃ -1 conjugate (ISIS 440762). These results demonstrate that the efficacy advantage of the GalNAc ₃ -1 conjugate is observed using antisense oligonucleotides with different chemically modified nucleosides and complementary to different targets, and in this example the modified nucleosides are limited ethyl sugar moieties (bicyclic sugar moieties).

Table 29

Effect of ASO Treatment on SRB-1 mRNA Levels in Balb/c Mice

Example 23: human peripheral blood mononuclear cell ( hPBMC ) assay protocol

hPBMC assay was performed using the BD Vautainer CPT tube method. Whole blood samples were obtained from donors who volunteered with informed consent at the US HealthWorks clinic (Faraday & El Camino Real, Carlsbad) and collected in 4-15 BD evacuator CPT 8 ml tubes (VWR Cat.# BD362753). . The approximate starting total blood volume in CPT tubes for each donor was recorded using the PBMC assay data sheet.

The tube was inverted 8-10 times to remix the blood sample just prior to centrifugation. ^{CPT tubes were centrifuged at room temperature (18-25 °} C) in a swing-out rotor at 15000-1800 RCF without brake (2700 RPM Beckman Allegra 6R) for 30 min. cells were recovered from the buffy coat interface (between the Ficoll and polymer gel layers); Transfer to sterile 50 ml conical tubes and pool up to 5 CPT tubes/50 ml conical tubes/donor. The cells were then washed twice with PBS (Ca++, no Mg++; GIBCO). The tube was filled to 50 ml and inverted several times to mix. The samples were then centrifuged at 330×g for 15 minutes at room temperature (1215 RPM on a Beckman Allegra 6R) and as much supernatant as possible was aspirated without touching the pellet. Vortex the tube to remove the cell pellet and resuspend the cells in RPMI+10% FBS+Pen/Strep (whole blood starting ˜1 ml/10 ml). A 60 μl sample was pipetted into a sample vial (Beckman Coulter) with 600 μl VersaLyse reagent (Beckman Coulter Cat# A09777) and vortexed gently for 10-15 seconds. Samples were incubated for 10 minutes at room temperature for 10 minutes, mixed again and counted. Cell suspensions were counted on a Vicell XR cell viability analyzer (Beckman Coulter) using PBMC cell types (a dilution factor of 1:11 was stored using different parameters). Viable cells/ml and viability were recorded. The cell suspension ^{was diluted to ml in 1×10 7} viable PBMC/RPMI+10% FBS+Pen/Strep.

Cells were plated at ^{5×10 5} in 50 μl/well of 96-well tissue culture plates (Falcon Microtest). 50 μl/well of 2x concentration oligo/control diluted in RPMI+10% FBS+pen/strep was added according to the experimental template (100 μl/well total). The plate was placed on a shaker and mixed for approximately 1 minute. After incubation at 37° C. for 24 hours; 5% CO ₂ , the plates were centrifuged at 400×g for 10 min and then the supernatant was removed for MSD cytokine assays (ie human IL-6, IL-10, IL-8 and MCP-1).

Example 24: GalNAc ₃ -One conjugated ASO for hPBMC in black pro-inflammatory evaluation of effectiveness

The pro-inflammatory effect in the hPBMC assay was evaluated using the protocol described in Example 23 for the antisense oligonucleotides (ASOs) listed in Table 30. ISIS 353512 is an internal standard known to be a high responder to IL-6 release in the assay. hPBMCs were isolated from fresh, supported donors and treated with ASO at concentrations of 0, 0.0128, 0.064, 0.32, 1.6, 8, 40 and 200 μM. After 24 hours of treatment, cytokine levels were measured.

The level of IL-6 was used as the primary readout. EC ₅₀ and E _max were calculated using standard procedures. Results are expressed as the average ratio of _{E max} /EC ₅₀ from the two donors and denoted as _{“E max} /EC _{50 ”.} A lower ratio indicates a relative decrease in the proinflammatory response and a higher ratio indicates a relative increase in the proinflammatory response.

Regarding the test compound, the lowest proinflammatory compound was PS/PO linked ASO (ISIS 616468). ISIS 647535, a GalNAc ₃ -1 conjugated ASO, was slightly less proinflammatory than its non-conjugated counterpart ISIS 304801. These results indicate that the incorporation of some PO linkages reduces the proinflammatory response and the addition of GalNAc ₃ -1 conjugates can make the compound more proinflammatory and reduce the proinflammatory response. Thus, one skilled in the art would know that antisense compounds comprising both mixed PS/PO linkages and GalNAc ₃ -1 conjugates produce a lower pro-inflammatory response compared to complete PS linked antisense compounds with or without GalNAc ₃ -1 conjugates. would expect to do These results show that the compounds having a GalNAc _3- 1 conjugate the antisense compounds, particularly reducing the PS content of less proinflammatory.

Taken together, these results suggest that _{GalNAc 3} -1 conjugated compounds, particularly compounds with reduced PS content, can be administered at higher doses than the corresponding intact PS antisense compound without GalNAc ₃ -1 conjugate. do. As the half-lives for these compounds are not expected to differ substantially, such higher dosing will result in less frequent dosing. In fact, such administration may be much less frequent, as the GalNAc ₃ -1 conjugated compound is more potent (see Examples 20-22) and requires re-administration if the concentration of the compound drops below the desired level based on potency. to be.

Table 30

Modified ASO

Subscript: Subscript: "e" represents a 2'-MOE modified nucleoside; "d" represents β-D-2'-deoxyribonucleoside;"k" represents a 6'- (S) -CH ₃ bicyclic nucleoside (eg cEt); "s" represents a phosphorothioate internucleoside linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO); And "o'" represents -OP(=O)(OH)-. The superscript "m" represents 5-methylcytosine. " A _do ' -GalNAc ₃ -1 _a " refers to a conjugate having the structure GalNAc ₃ - 1 shown in Example 9 attached to the 3'-end of the antisense oligonucleotide, as indicated.

Table 31

Pro-inflammatory effect of ASO targeting ApoC III in hPBMC assay

Example 25: in vitro at human ApoC targeting III GalNAc ₃ -One Konjuge Effect of modified ASO

ISIS 304801 and 647535 described above were tested in vitro. Primary hepatocyte cells from transgenic mice at a density of 25,000 cells per well were treated with modified oligonucleotides at concentrations of 0.03,0.08, 0.24, 0.74, 2.22, 6.67 and 20 μM. After a treatment period of approximately 16 hours, RNA was isolated from the cells and mRNA levels measured by quantitative real-time PCR and hApoC III mRNA levels adjusted according to total RNA content, as measured by ribogrin.

IC ₅₀ was calculated using standard methods and the results are presented in Table 32. As illustrated, comparable efficacy was observed in cells treated with ISIS 647535 compared to control ISIS 304801.

Table 32

Modified ASO Targeting Human ApoC III in Primary Hepatocytes

In this experiment, the efficacy benefits of a large GalNAc ₃ -1 konju aggregation observed in vivo was not observed in vitro. Subsequent free uptake experiments in primary hepatocytes in vitro did not show increased efficacy of oligonucleotides comprising various GalNAc conjugates compared to oligonucleotides without GalNAc conjugates (see Examples 60, 82, and 92). .

Example 26: ApoC III ASO Effect of PO/PS Linking on Activity

Human ApoC III transgenic mice were injected intraperitoneally with 25 mg/kg of ISIS 304801, or ISIS 616468 (all described above) or PBS-treated controls were injected once weekly for 2 weeks. The treatment group consisted of 3 animals and the control group consisted of 4 animals. Before treatment as well as after the last dose, blood was drawn from each mouse and plasma samples were analyzed. Mice were sacrificed 72 hours after the last dose.

Samples were collected and analyzed for ApoC III protein levels in the liver as described above (Example 20). Data from these analyzes are shown in Table 33 below.

These results show a decrease in potency of antisense compounds with PO/PS (ISIS 616468) in the wing compared to total PS (ISIS 304801).

Table 33

Effect of ASO Treatment on ApoC III Protein Levels in Human ApoC III Transgenic Mice

Example 27: compound 56

Compound 56 is commercially available from Glen Research, or can be prepared according to a published procedure reported by Shchepinov et al., Nucleic Acids Research, 1997, 25(22), 4447-4454.

Example 28: compound 60 manufacturing

Compound 4 was prepared according to the procedure exemplified in Example 2. Compound 57 is commercially available. Compound 60 was identified by structural analysis.

Compound 57 is considered representative and is limited as other monoprotected substituted or unsubstituted alkyl diols, including but not limited to those provided herein, which may be used to prepare phosphoramidite having a given composition. not intended

Example 29: compound 63 manufacturing

Compounds 61 and 62 are prepared using procedures analogous to those reported by Tober et al., Eur . J. Org . Chem ., 2013 , 3, 566-577; and Jiang et al., Tetrahedron, 2007, 63(19), 3982-3988.

Alternatively, compound 63 was combined with Kim et al., Synlett, 2003, 12, 1838-1840; and Kim et al. , prepared using procedures analogous to those reported in the scientific and patent literature by published PCT International Application, WO 2004063208. Example 30: Preparation of compound 63b

Compound 63a is prepared using procedures analogous to those reported by Hanessian et al., Canadian Journal of Chemistry, 1996 , 74(9), 1731-1737.

Example 31: compound Manufacture of 63d

Compound 63c was prepared using procedures analogous to those reported by Chen et al., Chinese Chemical Letters, 1998, 9(5), 451-453.

Example 32: compound 67 manufacturing

Compound 64 was prepared according to the procedure illustrated in Example 2. Compound 65 was prepared according to Or et al. , prepared using procedures similar to those reported by published PCT International Application, WO 2009/003009. The protecting groups used for compound 65 are meant to be representative and are not intended to be limited as other protecting groups, including, but not limited to, those set forth herein that may be used.

Example 33: compound 70 manufacturing

Compound 64 was prepared according to the procedure illustrated in Example 2. Compound 68 is commercially available. The protecting groups used for compound 68 are meant to be representative and are not intended to be limited as other protecting groups, including, but not limited to, those provided herein that may be used.

Example 34: compound Preparation of 75a

Compound 75 is prepared using procedures published by Shchepinov et al., Nucleic Acids Research, 1997, 25(22), 4447-4454.

Example 35: compound 79 manufacturing

Compound 76 was prepared using the procedure published by Shchepinov et al., Nucleic Acids Research, 1997, 25(22), 4447-4454.

Example 36: compound Preparation of 79a

Compound 77 is prepared according to the procedure demonstrated in Example 35.

Example 37: solid at the 5' end through the support phosphodiester connected GalNAc ₃ -2 a conjugate containing conjugated General method for preparing oligomeric compound 82 (Method I)

wherein GalNAc ₃ -2 has the formula:

Konju gate group GalNAc ₃ -2 GalNAc ₃ The cluster moiety (GalNAc ₃ -2 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. wherein GalNAc ₃ -2 _a has the formula:

VIMAD-linked oligomeric compound 79b was prepared using standard procedures for: automated DNA/RNA synthesis (Dupouy et al., Angew . Chem . Int . Ed., 2006, 45, 3623-3627) . Phosphoramidite compounds 56 and 60 were prepared according to the procedures exemplified in Examples 27 and 28, respectively. The phosphoramidites demonstrated are representative and other phosphoramidite buildings including, but not limited to, those provided herein that can be used to prepare oligomeric compounds having a phosphodiester linked conjugate group at the 5' end. It is not intended to be limited to blocks. The order and amount of phosphoramidite added to the solid support can be adjusted to prepare the oligomeric compounds described herein in any desired order and composition.

Example 38: 5 ' at the end phosphodiester connected GalNAc ₃ -2 a conjugate Alternative process for preparing oligomeric compound 82 comprising (Method II)

VIMAD-linked oligomeric compound 79b was prepared using standard procedures for: automated DNA/RNA synthesis (Dupouy et al., Angew . Chem . Int . Ed., 2006, 45, 3623-3627) . Compound 79, a GalNAc ₃ -2 cluster phosphoramidite, was prepared according to the procedure exemplified in Example 35. _{This alternative method allows one-step installation of phosphodiester linked GalNAc 3} -2 conjugates into oligomeric compounds at the final stage of the synthesis. The exemplified phosphoramidite is considered representative and as other phosphoramidite building blocks, including but not limited to those provided herein, that can be used to prepare oligomeric compounds having a phosphodiester conjugate at the 5' end. It is not intended to be limiting. The order and amount of phosphoramidite added to the solid support can be adjusted to prepare the oligomeric compounds described herein in any desired order and composition.

Example 39: solid support through at the 5' end (modified for 5' end attachment GalNAc ₃ -One) GalNAc ₃ -3 a conjugate General method for preparing oligomeric compound 83h comprising

Compound 18 was prepared according to the procedure illustrated in Example 4. Compounds 83a and 83b are commercially available. Oligomeric compound 83e comprising phosphodiester linked hexylamine was prepared using standard oligonucleotide synthesis procedures. Treatment of the protected oligomeric compound with aqueous ammonia gave a 5'-GalNAc ₃ -3 conjugated oligomeric compound (83h).

wherein GalNAc ₃ -3 has the formula:

.

.

Konju GalNAc ₃ of the gate group ₃ GalNAc -3 The cluster moiety (GalNAc ₃ -3 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. wherein GalNAc ₃ -3 _a has the formula:

.

.

Example 40: solid support through at the 3' end phosphodiester connected GalNAc ₃ -4 a conjugate General method for preparing oligomeric compound 89 comprising

wherein GalNAc ₃ -4 has the formula:

wherein CM is a cleavable moiety. In certain embodiments, the cleavable moiety is:

Konju gate group GalNAc ₃ -4 GalNAc ₃ Cluster moieties (GalNAc ₃ -4 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. wherein GalNAc ₃ -4 _a has the formula:

The protected Unylinker functionalized solid support compound 30 is commercially available. Compound 84 is prepared using procedures analogous to those reported by literature: (hchepinov et al., Nucleic Acids Research, 1997, 25(22) , 4447-4454; Shchepinov et al., Nucleic Acids Research, 1999, 27 , 3035-3041; and Hornet et al., Nucleic Acids Research, 1997, 25 , 4842-4849).

The phosphoramidite building blocks, compounds 60 and 79a, were prepared according to the procedures exemplified in Examples 28 and 36. The exemplified phosphoramidite is considered representative and is intended to be limited as other phosphoramidite building blocks that may be used to prepare oligomeric compounds having a phosphodiester linked conjugate at the 3' end in the desired order and composition. doesn't happen The order and amount of phosphoramidite added to the solid support can be adjusted to prepare the oligomeric compounds described herein in any desired order and composition.

Example 41: solid phase technology through at the 5' position phosphodiester connected GalNAc ₃ -2 ( Example 37 note, Bx adenine) a conjugate containing ASO's General manufacturing method (manufacturing of ISIS 661134)

Unless otherwise stated, all reagents and solutions used for the synthesis of oligomeric compounds are purchased from commercial sources. Standard phosphoramidite building blocks and solid supports are used for incorporation of nucleoside residues including, for example, T, A, G, and ^{m C residues.} The phosphoramidite compounds 56 and 60 used were used to synthesize _{a GalNAc 3} -2 conjugate phosphodiester linked at the 5' end. A 0.1 M solution of phosphoramidite in anhydrous acetonitrile was used for β-D-2'-deoxyribonucleoside and 2'-MOE.

올리고파일럿 합성기 (40-200 μmol 크기) 상에서 수행하였다. 커플링 단계의 경우, 포스포르아미다이트를 고체 지지체의 초기 로딩에 대해 4배 과량으로 전달하였고, 포스포르아미다이트 커플링을 10분 동안 수행하였다. 모든 다른 단계는 제조자에 의해 공급된 표준 프로토콜을 따랐다. 톨루엔 중의 6% 디클로로아세트산 용액을 뉴클레오티드의 5'-하이드록실 그룹으로부터 디메톡시트리틸 (DMT) 그룹을 제거하는데 사용하였다. 무수 CH₃CN 중 4,5-디시아노이미다졸 (0.7 M)은 커플링 단계 동안에 활성제로서 사용되었다. 포스포로티오에이트 연결을 3분의 접촉 시간 동안에 1:1 피리딘/CH3CN 중의 크산탄 하이드라이드의 0.1 M 용액을 이용한 황화에 의해 도입하였다. 6% 물을 함유하는 CH3CN 중의 20% tert-부틸하이드로퍼옥사이드의 용액을 산화제로서 사용하여 12분의 접촉 시간으로 포스포디에스테르 뉴클레오시드간 연결을 제공하였다. ASO synthesis was performed on ABI 394 synthesizer (1-2 μmol size) or GE Healthcare Bioscience by phosphoramidite coupling method on VIMAD solid support (110 μmol/g, Guzaev et al., 2003) packed in column.

performed on an oligopilot synthesizer (40-200 μmol size). For the coupling step, phosphoramidite was delivered in a 4-fold excess relative to the initial loading of the solid support, and phosphoramidite coupling was performed for 10 minutes. All other steps followed the standard protocol supplied by the manufacturer. A solution of 6% dichloroacetic acid in toluene was used to remove the dimethoxytrityl (DMT) group from the 5'-hydroxyl group of the nucleotide. 4,5-dicyanoimidazole (0.7 M) in anhydrous CH ₃ CN was used as activator during the coupling step. Phosphorothioate linkages were introduced by sulfiding with a 0.1 M solution of xanthan hydride in 1:1 pyridine/CHCN for a contact time of 3 minutes. A solution of 20% tert-butylhydroperoxide in CH3CN containing 6% water was used as the oxidizing agent to provide phosphodiester internucleoside linkages with a contact time of 12 minutes.

After the desired sequence was assembled, the cyanoethyl phosphate protecting group was deprotected using 20% diethylamine in toluene (v/v) with a contact time of 45 min. The solid-support bound ASO was suspended in aqueous ammonia (28-30 wt %) and heated at 55° C. for 6 hours.

Then, the unbound ASO was filtered off and the ammonia was removed by heating. The residue was purified by high pressure liquid chromatography on a strong anion exchange column (GE Healthcare Bioscience, Source 30Q, 30 μm, 2.54 x 8 cm, A = 100 mM ammonium acetate in 30% aqueous CHCN, B = 1.5 M in A. NaBr, 0-40% B at 60 min, flow rate 14 mL min-1, λ = 260 nm). The residue was desalted by HPLC on a reverse phase column to give the desired ASO in isolated yields of 15-30% based on the initial loading on the solid support. ASO was characterized by ion-pair-HPLC coupled MS analysis with an Agilent 1100 MSD system.

Table 34

SRB - 1 targeting at the 5' position phosphodiester connected GalNAc ₃ -2 a conjugate containing ASO

Subscript: Subscript: "e" represents a 2'-MOE modified nucleoside; "d" represents β-D-2'-deoxyribonucleoside;"k" represents a 6'- (S) -CH ₃ bicyclic nucleoside (eg cEt); "s" represents a phosphorothioate internucleoside linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO); And "o'" represents -OP(=O)(OH)-. The superscript "m" represents 5-methylcytosine. The structure of "GalNAc ₃ -2 _a " is shown in Example 37.

Example 42: solid phase 5' position via technology GalNAc ₃ -3 a conjugate containing ASO's General manufacturing method (manufacturing of ISIS 661166)

Synthesis for ISIS 661166 was performed using procedures similar to those exemplified in Examples 39 and 41.

ISIS 661166 is a 5-10-5 MOE gapmer, wherein the 5' position contains a GalNAc ₃ -3 conjugate. ASO was characterized by ion-pair-HPLC coupled MS analysis with an Agilent 1100 MSD system.

Table 34a

Malat -1 targeting hexylamino phosphodiester at the 5' position through the connection GalNAc ₃ -3 a conjugate containing ASO

Subscript: “e” indicates 2′-MOE modified nucleoside; “d” indicates β-D-2′-deoxyribonucleoside; “s” indicates phosphorothioate nucleoside represents a cross linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO); and "o'" represents -OP(=O)(OH)-. Superscript "m" represents 5-methylcytosine _{The structure of "5'-GalNAc 3} -3a" is shown in Example 39.

Example 43: in vivo at SRB -1 targeting at the 5' end phosphodiester connected GalNAc ₃ -2 ( Example See 37 and 41; Bx adenine) in a dose-dependent study

_{ISIS 661134 (see Example 41) comprising a GalNAc 3} -2 conjugate phosphodiester linked at the 5' end was tested in a dose-dependent study for antisense inhibition of SRB-1 in mice. Unconjugated ISIS 440762 and 651900 (GalNAc ₃ -1 conjugate at the 3' end, see Example 9) were included in the study for comparison and are previously described in Table 17.

cure

6-week-old male Balb/c mice (Jackson Laboratory, Bar Harbor, ME) were injected once subcutaneously at the doses given below as ISIS 440762, 651900, 661134 or PBS-treated controls. Each treatment group consisted of 4 animals. Mice were sacrificed 72 hours after final dosing and liver SRB-1 mRNA levels were measured using real-time PCR and RiboGreen® RNA quantification reagent according to standard protocols (Molecular Probes, Inc. Eugene, OR). SRB-1 mRNA levels were determined for total RNA (using ribogreen) prior to normalization to PBS-treated controls. Results below are presented as average percent of SRB-1 mRNA levels for each treatment group, normalized to PBS-treated controls, and are expressed as “% PBS”. The ED ₅₀ was determined using a method similar to that previously described and is provided below.

As illustrated in Table 35, antisense oligonucleotide treatment lowered SRB-1 mRNA levels in a dose-dependent manner. _{In fact, an antisense oligonucleotide comprising a GalNAc 3} -2 conjugate linked to the 5' end _{(ISIS 661134) or an antisense oligonucleotide comprising a GalNAc 3} -1 conjugate linked to the 3' end (ISIS 651900) is a beacon It showed substantial efficacy improvement compared to the conjugated antisense oligonucleotide (ISIS 440762). _{In addition, ISIS 661134 comprising a GalNAc 3} -2 conjugate linked to a phosphodiester at the 5' end was equivalent in efficacy to ISIS 651900 comprising a _{GalNAc 3} -1 conjugate at the 3' end.

Table 35

SRB targeting -1 GalNAc ₃ -1 or GalNAc ₃ containing -2 ASO

The _{structures for 3'GalNAc 3} -1 and 5'GalNAc ₃ -2 were previously described in Examples 9 and 37.

Pharmacokinetic analysis (PK)

The PK of ASO from the high dose group (7 mg/kg) was investigated and evaluated in the same manner as exemplified in Example 20. Liver samples were minced and extracted using standard protocols. Full-length metabolites of 661134 (5' GalNAc ₃ -2) and ISIS 651900 (3' GalNAc ₃ -1) were identified and their masses were determined by high-resolution mass spectrometry analysis. The above results showed that the major metabolite detected for ASO (ISIS 661134) comprising a _{GalNAc 3} -2 conjugate linked to the 5' end was ISIS 440762 (data not shown). No additional metabolites were observed at detectable levels. In contrast to its counterpart, an additional metabolite similar to that previously reported in Table 23a was observed for ASO with a _{GalNAc 3 -1 conjugate at the 3' end (ISIS 651900).} These results suggest that having phosphodiester linked GalNAc ₃ -1 or GalNAc ₃ -2 conjugates can improve the PK profile of ASO without abrogating its efficacy.

Example 44: SRB -1 targeting at the 3' end GalNAc ₃ -One a conjugate containing ASO (see Example 9) of antisense Effect of PO/PS Linkage on Inhibition

_{ISIS 655861 and 655862, each containing a GalNAc 3} -1 conjugate at the 3' end targeting SRB-1, were tested for their ability to inhibit SRB-1 in mice in a single dose study. The parent unconjugated compound, ISIS 353382, was included in the study for comparison.

ASO is a 5-10-5 MOE gapmer, wherein the gap region contains 10 2'-deoxyribonucleosides and each wing region contains 5 2'-MOE modified nucleosides. ASO was prepared using a method similar to that previously exemplified in Example 19 and is described in Table 36 below.

Table 36

SRB - 1 targeting at the 3' end GalNAc ₃ -One a conjugate modified to contain ASO

Subscript: “e” indicates 2′-MOE modified nucleoside; “d” indicates β-D-2′-deoxyribonucleoside; “s” indicates phosphorothioate nucleoside represents a cross linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO); and "o'" represents -OP(=O)(OH)-. Superscript "m" represents 5-methylcytosine _{The structure of "GalNAc 3} -1" is shown in Example 9.

cure

6-week-old male Balb/c mice (Jackson Laboratory, Bar Harbor, ME) were injected once subcutaneously at the doses given below as ISIS 353382, 655861, 655862 or PBS-treated controls. Each treatment group consisted of 4 animals. Before treatment as well as after the last dose, blood was drawn from each mouse and plasma samples were analyzed. Mice were sacrificed 72 hours after final dosing and liver SRB-1 mRNA levels were measured using real-time PCR and RiboGreen® RNA quantification reagent according to standard protocols (Molecular Probes, Inc. Eugene, OR). SRB-1 mRNA levels were determined for total RNA (using ribogreen) prior to normalization to PBS-treated controls. Results below are presented as average percent of SRB-1 mRNA levels for each treatment group, normalized to PBS-treated controls, and are expressed as “% PBS”. The ED ₅₀ was determined using a method similar to that described previously and is reported below.

As exemplified in Table 37, antisense oligonucleotide treatment lowered SRB-1 mRNA levels in a dose-dependent manner compared to PBS treated controls. _{In fact, antisense oligonucleotides comprising a GalNAc 3} -1 conjugate at the 3' end (ISIS 655861 and 655862) showed substantial efficacy improvement compared to unconjugated antisense oligonucleotides (ISIS 353382). Moreover, ISIS 655862 with mixed PS/PO linkages showed efficacy improvement compared to full PS (ISIS 655861).

Table 37

SRB - 1 targeting at the 3' end GalNAc ₃ -One a conjugate containing ASO's antisense Effect of PO/PS Linkage on Inhibition

Serum hepatic transaminase levels, alanine transaminase (ALT) and aspartate transaminase (AST) were measured for saline infused mice using standard protocols. Organ weights were also assessed. The results demonstrated that no elevations in transaminase levels (Table 38) or organ weights (data not shown) were observed in ASO treated mice compared to PBS controls (data not shown). Moreover, ASO with mixed PS/PO linkages (ISIS 655862) showed similar transaminase levels compared to total PS (ISIS 655861).

Table 38

SRB - 1 targeting at the 3' end GalNAc ₃ -One a conjugate containing ASO's trans Effect of PO/PS Linkage on Suaminase Levels

Example 45: PFP Preparation of ester, compound 110a

Compound 4 (9.5 g, 28.8 mmol) was treated individually with compound 103a or 103b (38 mmol), and molecular sieves with TMSOTf (0.5 eq.) in dichloromethane (200 mL) and stirred at room temperature for 16 h. At this time, the organic layer was filtered through celite and then washed with sodium bicarbonate, water and brine. The organic layer was then separated, dried over sodium sulfate, filtered and reduced under reduced pressure. The obtained oil was purified by silica gel chromatography (2%-->10% methanol/dichloromethane) to obtain compounds 104a and 104b in >80% yield. LCMS and proton NMR were consistent with the above structure.

Compounds 104a and 104b were treated under the same conditions as those of compounds 100a-d (Example 47) to obtain compounds 105a and 105b in >90% yield. LCMS and proton NMR were consistent with the above structure.

Compounds 105a and 105b were individually treated with compound 90 under the same conditions as compounds 901a-d to obtain compounds 106a (80%) and 106b (20%). LCMS and proton NMR were consistent with the above structure.

Compounds 106a and 106b were treated under the same conditions as compounds 96a-d (Example 47) to obtain 107a (60%) and 107b (20%). LCMS and proton NMR were consistent with the above structure.

Compounds 107a and 107b were treated under the same conditions as compounds 97a-d (Example 47) to obtain compounds 108a and 108b in 40-60% yield. LCMS and proton NMR were consistent with the above structure.

Compounds 108a (60%) and 108b (40%) were treated under the same conditions as compounds 100a-d (Example 47) to obtain compounds 109a and 109b in >80% yield. LCMS and proton NMR were consistent with the above structure.

Compound 109a was treated under the same conditions as compounds 101a-d (Example 47) to obtain compound 110a in 30-60% yield. LCMS and proton NMR were consistent with the above structure. Alternatively, compound 110b can be prepared in a similar manner as disclosed starting with compound 109b.

Example 46: PFP Esters (oligonucleotides) 111) and conjugation general procedure for; ISIS 666881 ( GalNAc ₃ - 10) of Produce

5'-hexylamino modified oligonucleotides were synthesized and purified using standard solid phase oligonucleotide procedures. The 5'-hexylamino modified oligonucleotide was dissolved in 0.1 M sodium tetraborate, pH 8.5 (200 μL) and 3 equivalents of the selected PFP esterified GalNAc ₃ clusters dissolved in DMSO (50 μL) were added. If the PFP ester precipitated upon addition of the ASO solution, DMSO was added until all the PFP ester was in solution. The reaction was completed after mixing for about 16 hours at room temperature. The resulting solution was diluted with 12 mL of water and then spun down at 3000 rpm on a spin filter with a mass cutoff of 3000 Da. This process was repeated twice to remove small molecule impurities. The solution was then lyophilized, redissolved in concentrated aqueous ammonia, mixed at room temperature for 2.5 hours, and then concentrated in vacuo to remove most of the ammonia. The conjugated oligonucleotides were purified, desalted by RP-HPLC and lyophilized _{to provide GalNAc 3} conjugated oligonucleotides.

Oligonucleotide 111 is gated konju and GalNAc ₃ -10. Konju gate group GalNAc ₃ -10 GalNAc ₃ Cluster portion (GalNAc ₃ -10 _a) is combined with any cuttable moiety can provide a wide range of conjugate groups. In certain embodiments, the cutting can moiety to an oligonucleotide synthesized using GalNAc ₃ -10 nucleotides -P (= O) as shown in (ISIS 666881) (OH) -A d -P (= O) (OH )-to be. The structure of GalNAc ₃ -10 (GalNAc ₃ -10 _a -CM-) is as follows:

ISIS 666881 was prepared according to the general procedure. 5'-hexylamino modified oligonucleotide ISIS 660254 was synthesized and purified using standard solid phase oligonucleotide procedures. ISIS 660254 (40 mg, 5.2 μmol) was dissolved in 0.1 M sodium tetraborate, pH 8.5 (200 μL) and 3 equivalents of PFP ester (compound 110a) dissolved in DMSO (50 μL) was added. The PFP ester precipitated upon addition to the ASO solution requiring additional DMSO (600 μL) to completely dissolve the PFP ester. The reaction was completed after mixing at room temperature for 16 hours. The solution was diluted with water to a total volume of 12 mL and spun down at 3000 rpm on a spin filter with a mass cutoff of 3000 Da. This process was repeated twice to remove small molecule impurities. The solution was lyophilized, redissolved in concentrated aqueous ammonia while mixing at room temperature for 2.5 hours, and then concentrated in vacuo to remove most of the ammonia. The conjugated oligonucleotide was purified, desalted by RP-HPLC, and lyophilized to obtain ISIS 666881 in a yield of 90% by weight (42 mg, 4.7 μmol).

GalNAc ₃ -10 conjugated oligonucleotide

Capital letters represent the nucleobase for each nucleoside and ^m C represents 5-methyl cytosine. subscript: "e" represents a 2'-MOE modified nucleoside; "d" represents β-D-2'-deoxyribonucleoside;"s" represents a phosphorothioate internucleoside linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO); And "o'" represents -OP(=O)(OH)-. Conjugate groups are in bold type.

Example 47: GalNAc ₃ - 8 Preparation of oligonucleotide 102 comprising

Triacid 90 (4 g, 14.43 mmol) was dissolved in DMF (120 mL) and N , N -diisopropylethylamine (12.35 mL, 72 mmol). Pentafluorophenyl trifluoroacetate (8.9 mL, 52 mmol) was added dropwise under argon, and the reaction was stirred at room temperature for 30 minutes. Boc-diamine 91a or 91b (68.87 mmol) was added along with N , N -diisopropylethylamine (12.35 mL, 72 mmol) and the reaction was stirred at room temperature for 16 h. At this time, DMF was reduced under reduced pressure >75% and the mixture was dissolved in dichloromethane. The organic layer was washed with sodium bicarbonate, water and brine. The organic layer was then separated, dried over sodium sulfate, filtered and reduced to oil under reduced pressure. The obtained oil was purified by silica gel chromatography (2%-->10% methanol/dichloromethane) to give compounds 92a and 92b in approximately 80% yield. LCMS and proton NMR were consistent with the above structure.

Compound 92a or 92b (6.7 mmol) was treated with 20 mL of dichloromethane and 20 mL of trifluoroacetic acid at room temperature for 16 hours. The resulting solution was evaporated, dissolved in methanol, and treated with DOWEX-OH resin for 30 minutes. The obtained solution was filtered and reduced under reduced oil pressure to obtain compounds 93a and 93b in 85-90% yield.

Compound 7 or 64 (9.6 mmol) was treated with HBTU (3.7g, 9.6 mmol) and N , N -diisopropylethylamine (5 mL) in DMF (20 mL) for 15 min. To this was added either compound 93a or 93b (3 mmol) and stirred at room temperature for 16 hours. At this time, DMF was reduced under reduced pressure >75% and the mixture was dissolved in dichloromethane. The organic layer was washed with sodium bicarbonate, water and brine. The organic layer was then separated, dried over sodium sulfate, filtered and reduced to oil under reduced pressure. The obtained oil was purified by silica gel chromatography (5%-->20% methanol/dichloromethane) to obtain compounds 96a-d in 20-40% yield. LCMS and proton NMR were consistent with the above structure.

Compounds 96a-d (0.75 mmol) were individually hydrogenated on Raney nickel in ethanol (75 mL) for 3 h. At this time, the catalyst was removed by filtration through celite, and ethanol was removed under reduced pressure to obtain compounds 97a-d in 80-90% yield. LCMS and proton NMR were consistent with the above structure.

Compound 23 (0.32 g, 0.53 mmol) was treated with HBTU (0.2 g, 0.53 mmol) and N , N -diisopropylethylamine (0.19 mL, 1.14 mmol) in DMF (30 mL) for 15 min. To this was added separately compounds 97a-d (0.38 mmol) and stirred at room temperature for 16 hours. At this time, DMF was reduced under reduced pressure >75% and the mixture was dissolved in dichloromethane. The organic layer was washed with sodium bicarbonate, water and brine. The organic layer was then separated, dried over sodium sulfate, filtered and reduced to oil under reduced pressure. The obtained oil was purified by silica gel chromatography (2%-->20% methanol/dichloromethane) to give compounds 98a-d in 30-40% yield. LCMS and proton NMR were consistent with the above structure.

Compound 99 (0.17 g, 0.76 mmol) was treated with HBTU (0.29 g, 0.76 mmol) and N , N -diisopropylethylamine (0.35 mL, 2.0 mmol) in DMF (50 mL) for 15 min. To this was separately added compounds 97a-d (0.51 mmol) and stirred at room temperature for 16 hours. At this time, DMF was reduced under reduced pressure >75% and the mixture was dissolved in dichloromethane. The organic layer was washed with sodium bicarbonate, water and brine. The organic layer was then separated, dried over sodium sulfate, filtered and reduced to oil under reduced pressure. The obtained oil was purified by silica gel chromatography (5%-->20% methanol/dichloromethane) to obtain compounds 100a-d in 40-60% yield. LCMS and proton NMR were consistent with the above structure.

Compounds 100a-d (0.16 mmol) were individually hydrogenated against 10% Pd(OH)2/C in methanol/ethyl acetate (1:1, 50 mL) for 3 h. At this time, the catalyst was removed by filtration through celite, and the organics were removed under reduced pressure to obtain compounds 101a-d in 80-90% yield. LCMS and proton NMR were consistent with the above structure.

Compounds 101a-d (0.15 mmol) were individually dissolved in DMF (15 mL) and pyridine (0.016 mL, 0.2 mmol). Pentafluorophenyl trifluoroacetate (0.034 mL, 0.2 mmol) was added dropwise under argon, and the reaction was stirred at room temperature for 30 minutes. At this time, DMF was reduced under reduced pressure >75% and the mixture was dissolved in dichloromethane. The organic layer was washed with sodium bicarbonate, water and brine. The organic layer was then separated, dried over sodium sulfate, filtered and reduced to oil under reduced pressure. The obtained oil was purified by silica gel chromatography (2%-->5% methanol/dichloromethane) to give compounds 102a-d in about 80% yield. LCMS and proton NMR were consistent with the above structure.

An oligomeric compound 102 comprising a GalNAc ₃ -8 conjugate group was prepared using the general procedure exemplified in Example 46. The GalNAc ₃ cluster portion of the conjugate group GalNAc _{3 -8 (GalNAc 3} -8 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. In a preferred embodiment, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-.

The structure of GalNAc ₃ -8 (GalNAc ₃ -8 _a -CM-) is as follows:

.

.

Example 48: GalNAc ₃ - 7 Preparation of oligonucleotide 119 comprising

Compound 112 was synthesized according to the procedure described in the literature: ( J. Med . Chem . 2004, 47 , 5798-5808).

Compound 112 (5 g, 8.6 mmol) was dissolved in 1:1 methanol/ethyl acetate (22 mL/22 mL). Palladium hydroxide on carbon (0.5 g) was added. The reaction mixture was stirred at room temperature under hydrogen for 12 h. The reaction mixture was filtered through a pad of Celite and washed with the pad with 1:1 methanol/ethyl acetate. The filtrate and washings were combined and concentrated to dryness to obtain compound 105a (quantitative). The structure was confirmed by LCMS.

Compound 113 (1.25 g, 2.7 mmol), HBTU (3.2 g, 8.4 mmol) and DIEA (2.8 mL, 16.2 mmol) were dissolved in anhydrous DMF (17 mL) and the reaction mixture was stirred at room temperature for 5 minutes. To this was added a solution of compound 105a (3.77 g, 8.4 mmol) in anhydrous DMF (20 mL). The reaction was stirred at room temperature for 6 hours. The solvent was removed under reduced pressure to give an oil. The residue was dissolved in CH ₂ Cl ₂ (100 mL) and washed with aqueous saturated NaHCO ₃ solution (100 mL) and brine (100 mL). The organic phase was separated, dried (Na ₂ SO ₄ ), filtered and evaporated. The residue was purified by silica gel column chromatography and eluted with 10-20 % MeOH in dichloromethane to give compound 114 (1.45 g, 30%). The structure ^{was confirmed by LCMS and 1} H NMR analysis.

Compound 114 (1.43 g, 0.8 mmol) was dissolved in 1:1 methanol/ethyl acetate (4 mL/4 mL). Palladium on carbon (0.14 g wet) was added. The reaction mixture was washed with hydrogen and stirred at room temperature under hydrogen for 12 hours. The reaction mixture was filtered through a pad of Celite. The Celite pad was washed with methanol/ethyl acetate (1:1). The filtrate and washings were combined together and evaporated under reduced pressure to give compound 115 (quantitative). The structure ^{was confirmed by LCMS and 1} H NMR analysis.

Compound 83a (0.17 g, 0.75 mmol), HBTU (0.31 g, 0.83 mmol) and DIEA (0.26 mL, 1.5 mmol) were dissolved in anhydrous DMF (5 mL) and the reaction mixture was stirred at room temperature for 5 minutes. To this was added a solution of compound 115 (1.22 g, 0.75 mmol) in anhydrous DMF and the reaction was stirred at room temperature for 6 hours. The solvent was removed under reduced pressure and the residue was dissolved in _{CH 2} Cl _{2 .} The organic layer was washed with aqueous saturated NaHCO ₃ solution and brine, _{dried over anhydrous Na 2} SO ₄ and filtered. The organic layer was concentrated to dryness, and the obtained residue was purified by silica gel column chromatography and eluted with 3-15% MeOH in dichloromethane to give compound 116 (0.84 g, 61%). The structure was confirmed by LC MS and ¹ H NMR analysis.

Compound 116 (0.74 g, 0.4 mmol) was dissolved in 1:1 methanol/ethyl acetate (5 mL/5 mL). Palladium on carbon (wet, 0.074 g) was added. The reaction mixture was washed with hydrogen and stirred at room temperature under hydrogen for 12 hours. The reaction mixture was filtered through a pad of Celite. The Celite pad was washed with methanol/ethyl acetate (1:1). The filtrate and washings were combined together and evaporated under reduced pressure to give compound 117 (0.73 g, 98%). The structure ^{was confirmed by LCMS and 1} H NMR analysis.

Compound 117 (0.63 g, 0.36 mmol) was dissolved in anhydrous DMF (3 mL). To this solution were added N , N -diisopropylethylamine (70 μL, 0.4 mmol) and pentafluorophenyl trifluoroacetate (72 μL, 0.42 mmol). The reaction mixture was stirred at room temperature for 12 h and poured into aqueous saturated NaHCO ₃ solution. The mixture was extracted with dichloromethane, washed with brine and dried over anhydrous Na ₂ SO ₄ . The dichloromethane solution was concentrated to dryness, purified by silica gel column chromatography, and eluted with 5-10% MeOH in dichloromethane to obtain compound 118 (0.51 g, 79%). The structure was confirmed by ^{LCMS and 1} H and ¹ H and ^{19 F NMR.}

Oligomeric compound 119 containing a GalNAc ₃ -7 conjugate group was prepared using the general procedure exemplified in Example 46. The GalNAc ₃ cluster portion of the conjugate group GalNAc _{3-7 (GalNAc 3-7 a} ) can be combined with any cleavable moiety to provide a variety of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-.

The structure of GalNAc ₃ -7 (GalNAc ₃ -7 _a -CM-) is as follows:

.

.

Example 49: GalNAc ₃ Preparation of oligonucleotide 132 comprising -5

Compound 120 (14.01 g, 40 mmol) and HBTU (14.06 g, 37 mmol) were dissolved in anhydrous DMF (80 mL). Triethylamine (11.2 mL, 80.35 mmol) was added and stirred for 5 min. The reaction mixture was cooled in an ice bath and a solution of compound 121 (10 g, mmol) in anhydrous DMF (20 mL) was added. Additional triethylamine (4.5 mL, 32.28 mmol) was added and the reaction mixture was stirred for 18 h under argon atmosphere. The reaction was observed by TLC (ethyl acetate:hexanes; 1:1; Rf = 0.47). The solvent was removed under reduced pressure. The residue was taken up in EtOAc (300 mL) and washed with 1M NaHSO ₄ (3×150 mL), aqueous saturated NaHCO ₃ solution (3×150 mL) and brine (2×100 mL). The organic layer was dried over Na ₂ SO ₄ . The drying agent was removed by filtration and the organic layer was concentrated by rotary evaporation. The crude mixture was purified by silica gel column chromatography and eluted with 35 50% EtOAc in hexanes to give compound 122 (15.50 g, 78.13%). The structure ^{was confirmed by LCMS and 1} H NMR analysis. Mass m/z 589.3 [M + H] ⁺ .

A solution of LiOH (92.15 mmol) in water (20 mL) and THF (10 mL) was added to a cooled solution of compound 122 (7.75 g, 13.16 mmol) in methanol (15 mL). The reaction mixture was stirred at room temperature for 45 min and observed by TLC (EtOAc:Hexanes; 1:1). The reaction mixture was concentrated to half volume under reduced pressure. The remaining solution was cooled in an ice bath and neutralized by addition of concentrated HCl. The reaction mixture was diluted, extracted with EtOAc (120 mL) and washed with brine (100 mL). An emulsion was formed and removed by standing overnight. The organic layer was separated _{, dried (Na 2} SO ₄ ), filtered and evaporated to give compound 123 (8.42 g). Residual salts are likely to be the cause of the excess mass. LCMS is consistent with the structure. The product was used without any further purification. MWcal:574.36; MWfd:575.3 [M + H] ⁺ .

Compound 126 was synthesized according to the procedure described in the literature: ( J. Am. Chem . Soc. 2011, 133 , 958-963).

Compound 123 (7.419 g, 12.91 mmol), HOBt (3.49 g, 25.82 mmol) and compound 126 (6.33 g, 16.14 mmol) were dissolved in DMF (40 mL) and the resulting reaction mixture was cooled in an ice bath. To this were added N , N -diisopropylethylamine (4.42 mL, 25.82 mmol), PyBop (8.7 g, 16.7 mmol) followed by Bop coupling reagent (1.17 g, 2.66 mmol) under argon atmosphere. The ice bath was removed and the solution warmed to room temperature. The reaction was complete after 1 h as determined by TLC (DCM:MeOH:AA; 89:10:1). The reaction mixture was concentrated under reduced pressure. The residue was dissolved in EtOAc (200 mL) and washed with 1 M NaHSO ₄ (3×100 mL), aqueous saturated NaHCO ₃ (3×100 mL) and brine (2×100 mL). The organic phase was separated, dried (Na ₂ SO ₄ ), filtered and concentrated. The residue was purified by silica gel column chromatography with a gradient of 50% hexanes/EtOAc to 100% EtOAc to give compound 127 (9.4 g) as a white foam. LCMS and ¹ H NMR were consistent with the structure. Mass m/z 778.4 [M + H] ⁺ .

Trifluoroacetic acid (12 mL) was added to a solution of compound 127 (1.57 g, 2.02 mmol) in dichloromethane (12 mL) and stirred at room temperature for 1 h. The reaction mixture was co-evaporated with toluene (30 mL) under reduced pressure to dryness. The obtained residue was co-evaporated twice with acetonitrile (30 mL) and toluene (40 mL) to give compound 128 (1.67 g) as trifluoro acetate salt which was used during the next step without further purification. LCMS and ¹ H NMR were consistent with the structure. Mass m/z 478.2 [M + H] ⁺ .

Compound 7 (0.43 g, 0.963 mmol), HATU (0.35 g, 0.91 mmol), and HOAt (0.035 g, 0.26 mmol) were combined together and _{dried in a round-bottom flask under reduced pressure over P 2} O ₅ for 4 hours and then dried over anhydrous It was dissolved in DMF (1 mL) and stirred for 5 min. To this was added a solution of compound 128 (0.20 g, 0.26 mmol) in anhydrous DMF (0.2 mL) and N , N-diisopropylethylamine (0.2 mL). The reaction mixture was stirred at room temperature under an argon atmosphere. The reaction was complete after 30 min as determined by LCMS and TLC (7% MeOH/DCM). The reaction mixture was concentrated under reduced pressure. The residue was dissolved in DCM (30 mL) and washed with 1 M NaHSO ₄ (3×20 mL), aqueous saturated NaHCO ₃ (3×20 mL) and brine (3×20 mL). The organic phase was separated, dried (Na ₂ SO ₄ ), filtered and concentrated. The residue was purified by silica gel column chromatography using 5-15% MeOH in dichloromethane to give compound 129 (96.6 mg). LC MS and 1 H NMR are consistent with the structure. Mass m/z 883.4 [M + 2H] ⁺ .

Compound 129 (0.09 g, 0.051 mmol) was dissolved in methanol (5 mL) in a 20 mL scintillation vial. A small amount of 10% Pd/C (0.015 mg) was added thereto, and the reaction vessel was washed _{with H 2 gas.} The reaction mixture was stirred at room temperature under H ₂ atmosphere for 18 h. The reaction mixture was filtered through a pad of Celite and the Celite pad was washed with methanol. The filtrate and washings were collected and concentrated under reduced pressure to obtain compound 130 (0.08 g). LCMS and ¹ H NMR were consistent with the structure. The product was used without further purification. Mass m/z 838.3 [M + 2H] ⁺ .

To a 10 mL marked round bottom flask was added compound 130 (75.8 mg, 0.046 mmol), 0.37 M pyridine/DMF (200 μL) and a stir bar. To this solution, 0.7 M pentafluorophenyl trifluoroacetate/DMF (100 μL) was added dropwise with stirring. The reaction was complete after 1 h as determined by LC MS. The solvent was removed under reduced pressure and the residue was dissolved in _{CHCl 3 (˜10 mL).} The organic layer _{was partitioned 3 times each against NaHSO 4} (1 M, 10 mL), aqueous saturated NaHCO ₃ (10 mL) and brine (10 mL). The organic phase was separated, _{dried over Na 2} SO ₄ , filtered and concentrated to give compound 131 (77.7 mg). LCMS is consistent with the structure. Used without further purification. Mass m/z 921.3 [M + 2H] ⁺ .

Oligomeric compound 132 comprising a GalNAc ₃ -5 conjugate group was prepared using the general procedure exemplified in Example 46. The GalNAc ₃ cluster portion of the conjugate group GalNAc _{3-5 a (GalNAc 3-5 a} ) can be combined with any cleavable moiety to provide a variety of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-.

The structure of GalNAc ₃ -5 (GalNAc ₃ -5 _a -CM-) is as follows:

.

.

Example 50: GalNAc ₄ - 11 Preparation of oligonucleotide 144 comprising

Synthesis of compound 134. To a Maryfield flask was added aminomethyl VIMAD resin (2.5 g, 450 μmol/g) washed with acetonitrile, dimethylformamide, dichloromethane and acetonitrile. The resin was expanded in acetonitrile (4 mL). Compound 133 was pre-activated in a 100 mL round bottom flask by addition of 20 (1.0 mmol, 0.747 g), TBTU (1.0 mmol, 0.321 g), acetonitrile (5 mL) and DIEA (3.0 mmol, 0.5 mL). The solution was stirred for 5 minutes and then added to the Maryfield flask with shaking. The suspension was stirred for 3 hours. The reaction mixture was drained and the resin was washed with acetonitrile, DMF and DCM. The fresh resin loading was quantified by measuring the absorbance of DMT cations at 500 nm (extinction coefficient = 76000) in DCM and was determined to be 238 μmol/g. The resin was suspended in acetic anhydride solution three times for 10 minutes and camped.

Solid support bound compound 141 was synthesized using an iterative Fmoc-based solid phase peptide synthesis method. A small amount of the solid support was withdrawn and suspended in aqueous ammonia (28-30 wt%) for 6 hours. The cleaved compound was analyzed by LC-MS and the observed mass was consistent with the structure. Mass m/z 1063.8 [M + 2H] ⁺ .

Solid support bound compound 142 was synthesized using a solid phase peptide synthesis method.

Solid support bound compound 143 was synthesized using standard solid phase synthesis on a DNA synthesizer.

The solid support-bound compound 143 was suspended in aqueous ammonia (28-30 wt%) and ^{heated at 55 °} C for 16 hours. The solution was cooled and the solid support was filtered. The filtrate was concentrated and the residue was dissolved in water and purified by HPLC on a strong anion exchange column. Fractions containing full length compound 144 were pooled and desalted. The obtained GalNAc ₄ -11 konju gated oligomeric compounds were analyzed by LC-MS, with the observed mass was consistent with the structure.

Konju gate group GalNAc ₄ -11 GalNAc ₄ Cluster portion (GalNAc ₄ -11 _a) is combined with any cuttable moiety can provide a wide range of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-.

The structure of _{GalNAc 4 -11 (GalNAc 4 -11 a} -CM) are as follows:

.

.

Example 51: GalNAc ₃ - 6 Preparation of oligonucleotide 155 comprising

Compound 146 was synthesized as described in literature: ( Analytical Biochemistry 1995, 229 , 54-60).

Compound 4 (15 g, 45.55 mmol) and compound 35b (14.3 g, 57 mmol) were dissolved in CH ₂ Cl ₂ (200 ml). Activated molecular sieves (4 A. 2 g, powdered) were added and the reaction stirred under nitrogen atmosphere for 30 minutes. TMS-OTf was added (4.1 ml, 22.77 mmol) and the reaction was stirred overnight at room temperature. Upon completion, the reaction was _{quenched by pouring it into a solution of saturated aqueous NaHCO 3} (500 ml) and crushed ice (~ 150 g). The organic layer was isolated, washed with brine, _{dried over MgSO 4} , filtered and concentrated under reduced pressure to give an orange oil. The crude material was purified by silica gel column chromatography _{and eluted with 2-10% MeOH in CH 2} Cl ₂ to give compound 112 (16.53 g, 63%). LCMS and ¹ HNMR were consistent with the expected compound.

Compound 112 (4.27 g, 7.35 mmol) was dissolved in 1:1 MeOH/EtOAc (40 ml). The reaction mixture was purged by bubbling the solution with a stream of argon for 15 minutes. Perlman's catalyst (palladium hydroxide on carbon, 400 mg) was added and the solution was bubbled with hydrogen gas for 30 minutes. Upon completion ( _{TLC 10% MeOH in CH 2} Cl ₂ , and LCMS), the catalyst was removed by filtration through a pad of Celite. The filtrate was concentrated by rotary evaporation and briefly dried under vacuum to give compound 105a (3.28 g). LCMS and 1 H NMR were consistent with the desired product.

Compound 147 (2.31 g, 11 mmol) was dissolved in anhydrous DMF (100 mL). N , N -diisopropylethylamine (DIEA, 3.9 mL, 22 mmol) was added followed by HBTU (4 g, 10.5 mmol). The reaction mixture was stirred under nitrogen for ˜15 min. To this, a solution of compound 105a (3.3 g, 7.4 mmol) in dry DMF was added and stirred under nitrogen atmosphere for 2 hours. The reaction was diluted with EtOAc and washed with saturated aqueous NaHCO ₃ and brine. The organic phase was separated, dried (MgSO ₄ ), filtered and concentrated to give an orange syrup. The crude material _{was purified by column chromatography in CH 2} Cl ₂ 2-5 % MeOH to give compound 148 (3.44 g, 73 %). LCMS and ¹ H NMR were consistent with the expected compound.

Compound 148 (3.3 g, 5.2 mmol) was dissolved in 1:1 MeOH/EtOAc (75 ml). The reaction mixture was purged by bubbling the solution with a stream of argon for 15 minutes. Perlman catalyst (palladium hydroxide on carbon) was added (350 mg). The solution was bubbled with hydrogen gas for 30 minutes. Upon completion (TLC 10% MeOH in DCM, and LCMS), the catalyst was removed by filtration through a pad of Celite. The filtrate was concentrated by rotary evaporation and dried briefly under high vacuum to give compound 149 (2.6 g). LCMS was consistent with the desired product. The residue was dissolved in dry DMF (10 ml) and it was used immediately in the next step.

Compound 146 (0.68 g, 1.73 mmol) was dissolved in dry DMF (20 ml). To this were added DIEA (450 μL, 2.6 mmol, 1.5 eq.) and HBTU (1.96 g, 0.5.2 mmol). The reaction mixture was stirred for 15 min at room temperature under nitrogen. A solution of compound 149 (2.6 g) in anhydrous DMF (10 mL) was added. The pH of the reaction was adjusted to pH = 9-10 by addition of DIEA (if necessary). The reaction was stirred at room temperature under nitrogen for 2 h. Upon completion, the reaction was diluted with EtOAc (100 mL) and washed with aq. sat. aq. NaHCO ₃ and brine. The organic phase was separated, dried (MgSO ₄ ), filtered and concentrated. The crude material was purified by silica gel column chromatography _{and eluted with 2-10% MeOH in CH 2} Cl ₂ to give compound 150 (0.62 g, 20 %). LCMS and ¹ H NMR were consistent with the expected compound.

Compound 150 (0.62 g) was dissolved in 1:1 MeOH/EtOAc (5 L). The reaction mixture was purged by bubbling the solution with a stream of argon for 15 minutes. Perlman catalyst (palladium hydroxide on carbon) was added (60 mg). The solution was bubbled with hydrogen gas for 30 minutes. Upon completion (TLC 10% MeOH in DCM, and LCMS), the catalyst was removed by filtration (syringe-tip Teflon filter, 0.45 μm). The filtrate was concentrated by rotary evaporation and dried briefly under high vacuum to give compound 151 (0.57 g). LCMS was consistent with the desired product. The product was dissolved in 4 mL dry DMF and used immediately in the next step.

Compound 83a (0.11 g, 0.33 mmol) was dissolved in anhydrous DMF (5 mL) and N , N -diisopropylethylamine (75 μL, 1 mmol) and PFP-TFA (90 μL, 0.76 mmol) were added. The reaction mixture turned magenta upon contact, then gradually turned orange over the next 30 minutes. The progress of the reaction was observed by TLC and LCMS. Upon completion (formation of PFP ester), a solution of compound 151 (0.57 g, 0.33 mmol) in DMF was added. The pH of the reaction was adjusted to pH = 9-10 by addition of N , N -diisopropylethylamine (if necessary). The reaction mixture was stirred under nitrogen for ~30 min. Upon completion, the majority of solvent was removed under reduced pressure. The residue _{was diluted with CH 2} Cl ₂ and washed with aqueous saturated NaHCO ₃ and brine. The organic phase was separated, _{dried over MgSO 4} , filtered and concentrated to give an orange syrup. The residue was purified by silica gel column chromatography ( _{2-10 % MeOH in CH 2} Cl ₂ ) to give compound 152 (0.35 g, 55 %). LCMS and ¹ H NMR were consistent with the expected compound.

Compound 152 (0.35 g, 0.182 mmol) was dissolved in 1:1 MeOH/EtOAc (10 mL). The reaction mixture was purged by bubbling the solution with a stream of argon for 15 minutes. Perlman catalyst (palladium hydroxide on carbon) was added (35 mg). The solution was bubbled with hydrogen gas for 30 minutes. Upon completion (TLC 10% MeOH in DCM, and LCMS), the catalyst was removed by filtration (syringe-tip Teflon filter, 0.45 μm). The filtrate was concentrated by rotary evaporation and briefly dried under high vacuum to give compound 153 (0.33 g, quantitative). LCMS was consistent with the desired product.

Compound 153 (0.33 g, 0.18 mmol) was dissolved in anhydrous DMF (5 mL) under nitrogen with stirring. To this were added N , N -diisopropylethylamine (65 μL, 0.37 mmol) and PFP-TFA (35 μL, 0.28 mmol). The reaction mixture was stirred under nitrogen for ~30 min. The reaction mixture turned magenta upon contact and gradually turned orange. More N ,-diisopropylethylamine was added to maintain the pH of the reaction mixture at pH=9-10. The progress of the reaction was observed by TLC and LCMS. Upon completion, the majority of solvent was removed under reduced pressure. The residue _{was diluted with CH 2} Cl ₂ (50 mL) and washed with aqueous saturated NaHCO ₃ and brine. The organic layer was _{dried over MgSO 4} , filtered and concentrated to give an orange syrup. The crude material was purified by column chromatography _{and eluted with 2-10% MeOH in CH 2} Cl ₂ to give compound 154 (0.29 g, 79 %). LCMS and ¹ H NMR were consistent with the expected compound.

The GalNAc ₃ -6 155 oligomeric compound comprising a conjugate group was prepared using the general procedure illustrated in Example 46. ₃ GalNAc cluster portion of the conjugate group _{GalNAc 3 -6 (GalNAc 3 -6 a} ) is combined with any cuttable moiety can provide a wide range of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-.

The structure of _{GalNAc 3 -6 (GalNAc 3 -6 a} -CM-) is as follows:

.

.

Example 52: GalNAc ₃ Preparation of oligonucleotide 160 comprising -9

Compound 156 was synthesized according to the procedure described in the literature: ( J. Med . Chem . 2004, 47 , 5798-5808).

Compound 156 (18.60 g, 29.28 mmol) was dissolved in methanol (200 mL). Palladium on carbon (6.15 g, 10 wt %, loading (dry basis), matrix carbon powder, wet) was added. The reaction mixture was stirred at room temperature under hydrogen for 18 h. The reaction mixture was filtered through a pad of Celite and the Celite pad was thoroughly rinsed with methanol. The combined filtrates were washed and concentrated to dryness. The residue was purified by silica gel column chromatography and eluted with 5-10 % methanol in dichloromethane to give compound 157 (14.26 g, 89%). Mass m/z 544.1 [MH] ^- .

Compound 157 (5 g, 9.17 mmol) was dissolved in anhydrous DMF (30 mL). HBTU (3.65 g, 9.61 mmol) and N , N -diisopropylethylamine (13.73 mL, 78.81 mmol) were added and the reaction mixture was stirred at room temperature for 5 minutes. To this was added a solution of compound 47 (2.96 g, 7.04 mmol). The reaction was stirred at room temperature for 8 hours. The reaction mixture was poured into saturated _{aqueous NaHCO 3} solution. The mixture was extracted with ethyl acetate and the organic layer was washed with brine, dried (over Na ₂ SO ₄ ), filtered and evaporated. The obtained residue was purified by silica gel column chromatography and eluted with 50% ethyl acetate in hexane to give compound 158 (8.25 g, 73.3%). The structure ^{was confirmed by MS and 1} H NMR analysis.

Compound 158 (7.2 g, 7.61 mmol) was dried _{under reduced pressure over P 2} O _{5 .} The dried compound was dissolved in anhydrous DMF (50 mL). Here, 1H-tetrazole (0.43 g, 6.09 mmol) and N-methylimidazole (0.3 mL, 3.81 mmol) and 2-cyanoethyl- N , N , N' , N' -tetraisopropyl phosphoro Diamidite (3.65 mL, 11.50 mmol) was added. The reaction mixture was stirred under argon atmosphere for 4 hours. The reaction mixture was diluted with ethyl acetate (200 mL). The reaction mixture was washed with _{saturated NaHCO 3 and brine.} The organic phase was separated, dried (Na ₂ SO ₄ ), filtered and evaporated. The residue was purified by silica gel column chromatography and eluted with 50-90 % ethyl acetate in hexane to give compound 159 (7.82 g, 80.5%). The structure ^{was confirmed by LCMS and 31} P NMR analysis.

Oligomeric compound 160 comprising a GalNAc ₃ -9 conjugate group was prepared using standard oligonucleotide synthesis procedures. Three units of compound 159 were coupled to a solid support followed by coupling to the nucleotide phosphoramidite. Treatment of the protected oligomeric compound with aqueous ammonia gave compound 160. Konju gate group GalNAc ₃ -9 GalNAc ₃ Cluster moieties (GalNAc ₃ -9 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-. The structure of GalNAc ₃ -9 (GalNAc ₃ -9 _a -CM) is as follows:

.

.

Example 53: compound 18 ( GalNAc ₃ -1a and GalNAc ₃ Alternative procedure for the preparation of -3a)

Lactone 161 was reacted with diamino propane (3-5 eq) or mono-Boc protected diamino propane (1 eq) to give alcohol 162a or 162b. When unprotected propanediamine was used in the reaction, excess diamine was removed by evaporation under high vacuum and the free amino group in 162a was protected using CbzCl to give 162b as a white solid after purification by column chromatography . Further reaction of alcohol 162b with compound 4 in the presence of TMSOTf gave 163a, which was converted to 163b by removal of the Cbz group using catalytic hydrogenation. Pentafluorophenyl (PFP) ester 164 was prepared by reacting triacid 113 (Reference Example 48) with PFPTFA (3.5 eq) and pyridine (3.5 eq) in DMF (0.1-0.5 M). The triester 164 was directly reacted with amine 163b (3-4 eq) and DIPEA (3-4 eq) to give compound 18. This method greatly facilitates the purification of the intermediate and minimizes the formation of by-products formed using the procedure described in Example 4.

Example 54: compound 18 ( GalNAc ₃ -1a and GalNAc ₃ Alternative procedure for the preparation of -3a)

triPFP ester 164 was prepared from acid 113 using the procedure outlined in Example 53 above and reacted with mono-Boc protected diamine to provide 165 in essentially quantitative yield. The Boc group was removed with hydrochloric acid or trifluoroacetic acid to give the triamine, which was reacted with PFP activated acid 166 in the presence of a suitable base such as DIPEA to provide compound 18.

PFP protected Gal-NAc acid 166 was prepared from the corresponding acid by treatment with PFPTFA (1-1.2 eq) and pyridine (1-1.2 eq) in DMF. The precursor acid was eventually prepared from the corresponding alcohol by oxidation with BAIB and TEMPO (0.2 eq) in acetonitrile and water. A sugar intermediate by reaction of the precursor alcohol with 1,6-hexanediol (or 1,5-pentanediol or other diol for other n values) (2-4 eq) and TMSOTf using the conditions previously described for Example 47 4 was prepared.

Example 55: in vivo at SRB 3' or 5'- to target -1 conjugate group (GalNAc ₃ Dose-dependent studies of oligonucleotides including -1, 3, 8 and 9)

The oligonucleotides listed below were tested in a dose-dependent study for antisense inhibition of SRB-1 in mice. The unconjugated ISIS 353382 is included as a standard. Each of the various GalNAc ₃ conjugate groups was attached at the 3' or 5' terminus of each oligonucleotide by a phosphodiester linked 2'-deoxyadenosine nucleoside (cleavable moiety).

Table 39

SRB Modified to target -1 ASO

Capital letters represent the nucleobase for each nucleoside and ^m C represents 5-methyl cytosine. subscript: "e" represents a 2'-MOE modified nucleoside; "d" represents β-D-2'-deoxyribonucleoside;"s" represents a phosphorothioate internucleoside linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO); And "o'" represents -OP(=O)(OH)-. Conjugate groups are in bold type.

The structure of "GalNAc ₃ -1 _a " is already shown in Example 9. The structure of “GalNAc ₃ -9” is shown in Example 52. The structure of “GalNAc ₃ -3” is shown in Example 39. The structure of “GalNAc ₃ -8” is shown in Example 47.

cure

6 week old male Balb/c mice (Jackson Laboratory, Bar Harbor, ME) were injected once subcutaneously with ISIS 353382, 655861, 664078, 661161, 665001 or saline at the doses shown below. Each treatment group consisted of 4 animals. Mice were sacrificed 72 hours after final administration and liver SRB-1 mRNA levels were measured using real-time PCR and RiboGreen® RNA quantification reagent according to standard protocols (Molecular Probes, Inc. Eugene, OR). The results below are presented as the average percentage of SRB-1 mRNA levels for each treatment group normalized to the saline control.

As demonstrated in Table 40, treatment with antisense oligonucleotides reduced SRB-1 mRNA levels in a dose-dependent manner. In fact, ₃ 'GalNAc 3 -1 and GalNAc linked phosphodiester at the terminal _3-9 conjugate (ISIS 655861 and ISIS 664078) and 5' GalNAc attached at the ends _3-3 and GalNAc ₃ -8 conjugate (ISIS 661161 and ISIS 665001) showed a substantial improvement in efficacy compared to the unconjugated antisense oligonucleotide (ISIS 353382). Moreover, _{ISIS 664078 with a GalNAc 3} −9 conjugate at the 3′ end was essentially equally potent compared to ISIS 655861 with a _{GalNAc 3 −1 conjugate at the 3′ end.} 5' conjugated antisense oligonucleotides, ISIS 661161 and ISIS 665001 comprising GalNAc ₃ -3 or GalNAc ₃ -9, respectively, were increased compared to 3' conjugated antisense oligonucleotides (ISIS 655861 and ISIS 664078). had efficacy.

Table 40

SRB targeting -1 GalNAc ₃ containing -1, 3, 8 or 9 ASO

Serum hepatic transaminase levels, alanine transaminase (ALT) and aspartate transaminase (AST) were measured for saline infused mice using standard protocols. Total bilirubin and BUN were also assessed. Changes in body weight were assessed without significant changes from the saline group. ALT, AST, total bilirubin and BUN values are shown in the table below.

Table 41

Example 56: in vivo at SRB 3' or 5'- to target -1 conjugate group (GalNAc ₃ -1, 2, 3, 5, 6, 7, and 10 comparisons of oligonucleotides, including

The oligonucleotides listed below were tested in a dose-dependent study for antisense inhibition of SRB-1 in mice. The unconjugated ISIS 353382 is included as a standard. Each of the various GalNAc ₃ conjugate groups was attached at the 5' end of each oligonucleotide by a phosphodiester linked 2'-deoxyadenosine nucleoside (cleavable moiety), with GalNAc _{3 attached only at the 3' end.} ISIS 655861 with conjugate groups is excluded.

Table 42

SRB Modified to target -1 ASO

Capital letters represent the nucleobase for each nucleoside and ^m C represents 5-methyl cytosine. subscript: "e" represents a 2'-MOE modified nucleoside; "d" represents β-D-2'-deoxyribonucleoside;"s" represents a phosphorothioate internucleoside linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO); And "o'" represents -OP(=O)(OH)-. Conjugate groups are in bold type.

The structure of "GalNAc ₃ -1 _a " is already shown in Example 9. The structure of "GalNAc ₃ -2 _a " is already shown in Example 37. The structure of "GalNAc ₃ -3 _a " is already shown in Example 39. The structure of "GalNAc ₃ -5 _a " is already shown in Example 49. The structure of “GalNAc ₃ -6 _a ” is already shown in Example 51. The structure of "GalNAc ₃ -7 _a " is already shown in Example 48. The structure of “GalNAc ₃ -10 _a ” is already shown in Example 46.

cure

6 week old male Balb/c mice (Jackson Laboratory, Bar Harbor, ME) were injected subcutaneously once with ISIS 353382, 655861, 664507, 661161, 666224, 666961, 666981, 666881 or saline at the doses shown below. Each treatment group consisted of 4 animals. Mice were sacrificed 72 hours after final administration and liver SRB-1 mRNA levels were measured using real-time PCR and RiboGreen® RNA quantification reagent according to standard protocols (Molecular Probes, Inc. Eugene, OR). The results below are presented as the average percentage of SRB-1 mRNA levels for each treatment group normalized to the saline control.

As demonstrated in Table 43, treatment with antisense oligonucleotides reduced SRB-1 mRNA levels in a dose-dependent manner. In fact, the conjugated antisense oligonucleotide showed a substantial improvement in efficacy compared to the unconjugated antisense oligonucleotide (ISIS 353382). 5' conjugated antisense oligonucleotides showed a slight increase in potency compared to 3' conjugated antisense oligonucleotides.

Table 43

Serum hepatic transaminase levels, alanine transaminase (ALT) and aspartate transaminase (AST) were measured for saline infused mice using standard protocols. Total bilirubin and BUN were also assessed. Changes in body weight were assessed without significant changes from the saline group. ALT, AST, total bilirubin and BUN values are shown in Table 44 below.

Table 44

Example 57: in vivo at ApoC 3'- to target III conjugate Duration of study of the action of oligonucleotides comprising groups

Mice were injected once at the doses specified below and monitored for ApoC-III and plasma triglyceride (plasma TG) levels over the course of 42 days. The study was performed using three transgenic mice expressing human APOC-III in each group.

Table 45

ApoC Modified targeting III ASO

Capital letters represent the nucleobase for each nucleoside and ^m C represents 5-methyl cytosine. subscript: "e" represents a 2'-MOE modified nucleoside; "d" represents β-D-2'-deoxyribonucleoside;"s" represents a phosphorothioate internucleoside linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO); And "o'" represents -OP(=O)(OH)-. Conjugate groups are in bold type.

The structure of "GalNAc ₃ -1 _a " is already shown in Example 9.

Table 46

ApoC III mRNA ( Day 1 brine % ) and plasma TG levels ( Day 1 brine % )

As can be seen from the table above, the duration of action was increased with the addition of the 3'-conjugate group compared to the unconjugated oligonucleotide. The duration of action was further increased for the conjugated mixed PO/PS oligonucleotide 647536 compared to the conjugated complete PS oligonucleotide 647535.

Example 58: in vivo at SRB 3'- to target -1 conjugate group ( GalNAc ₃ -1 and GalNAc ₄ −11 dose-dependent studies of oligonucleotides including

The oligonucleotides listed below were tested in a dose-dependent study for antisense inhibition of SRB-1 in mice. Unconjugated ISIS 440762 was included as an unconjugated standard. Each conjugate group was attached at the 3' end of each oligonucleotide by a phosphodiester linked 2'-deoxyadenosine nucleoside cleavable moiety.

The structure of "GalNAc ₃ -1 _a " is already shown in Example 9. The structure of “GalNAc ₃ -11 _a ” is already shown in Example 50.

cure

6 week old male Balb/c mice (Jackson Laboratory, Bar Harbor, ME) were injected once subcutaneously with ISIS 440762, 651900, 663748 or saline at the doses shown below. Each treatment group consisted of 4 animals. Mice were sacrificed 72 hours after final administration and liver SRB-1 mRNA levels were measured using real-time PCR and RiboGreen® RNA quantification reagent according to standard protocols (Molecular Probes, Inc. Eugene, OR). The results below are presented as the average percentage of SRB-1 mRNA levels for each treatment group normalized to the saline control.

As demonstrated in Table 47, treatment with antisense oligonucleotides reduced SRB-1 mRNA levels in a dose-dependent manner. 3 'terminal antisense oligonucleotides containing phosphodiester linked GalNAc ₃ -1 and GalNAc ₄ -11 conjugate in (ISIS 651900 and ISIS 663748) nucleotide is the substantial efficacy to raise the beacon main gate antisense compared to the nucleotide (ISIS 440762) showed improvement. The two conjugated oligonucleotides, GalNAc ₃ -1 and GalNAc ₄ -11, were equally potent.

Table 47

SRB Modified to target -1 ASO

Capital letters represent the nucleobase for each nucleoside and ^m C represents 5-methyl cytosine. subscript: "e" represents a 2'-MOE modified nucleoside; "k" represents a 6'- (S) -CH ₃ bicyclic nucleoside; "d" represents β-D-2'-deoxyribonucleoside;"s" represents a phosphorothioate internucleoside linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO); And "o'" represents -OP(=O)(OH)-. Conjugate groups are in bold type.

Serum hepatic transaminase levels, alanine transaminase (ALT) and aspartate transaminase (AST) were measured for saline infused mice using standard protocols. Total bilirubin and BUN were also assessed. Changes in body weight were assessed without significant changes from the saline group. ALT, AST, total bilirubin and BUN values are shown in Table 48 below.

Table 48

Example 59: in vivo FXI targeted GalNAc ₃ -One conjugated ASO's effect

The oligonucleotides listed below were tested in a multiple-dose study for antisense inhibition of FXI in mice. ISIS 404071 was included as an unconjugated standard. Each conjugate group was attached at the 3' end of each oligonucleotide by a phosphodiester linked 2'-deoxyadenosine nucleoside cleavable moiety.

Table 49

FXI modified to target ASO

Capital letters represent the nucleobase for each nucleoside and ^m C represents 5-methyl cytosine. subscript: "e" represents a 2'-MOE modified nucleoside; "d" represents β-D-2'-deoxyribonucleoside;"s" represents a phosphorothioate internucleoside linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO); And "o'" represents -OP(=O)(OH)-. Conjugate groups are in bold type.

The structure of "GalNAc ₃ -1 _a " is already shown in Example 9.

cure

6 week old male Balb/c mice (Jackson Laboratory, Bar Harbor, ME) were injected subcutaneously twice weekly for 3 weeks with ISIS 404071, 656172, 656173 or PBS treated controls at the doses shown below. Each treatment group consisted of 4 animals. Mice were sacrificed 72 hours after final dosing and liver FXI mRNA levels were measured using real-time PCR and RiboGreen® RNA quantitation reagent according to standard protocols (Molecular Probes, Inc. Eugene, OR). Plasma FXI protein levels were also measured using ELISA. FXI mRNA levels were determined for total RNA (using RiboGreen®) prior to normalization to PBS-treated controls. The results below are presented as the average percent of FXI mRNA levels for each treatment group. Data were normalized to PBS-treated controls and expressed as “% PBS”. The ED ₅₀ was determined using a method similar to that previously described and is provided below.

Table 50

factor XI mRNA (brine % )

As demonstrated in Table 50, treatment with antisense oligonucleotides reduced FXI mRNA levels in a dose-dependent manner. An oligonucleotide comprising a 3'-GalNAc ₃ -1 conjugate group showed a substantial improvement in efficacy compared to an unconjugated antisense oligonucleotide (ISIS 404071). An improvement in efficacy between the two conjugated oligonucleotides was additionally provided by replacing part of the PS linkage with PO (ISIS 656173).

As demonstrated in Table 50a, treatment with antisense oligonucleotides reduced FXI protein levels in a dose-dependent manner. An oligonucleotide comprising a 3'-GalNAc ₃ -1 conjugate group showed a substantial improvement in efficacy compared to an unconjugated antisense oligonucleotide (ISIS 404071). An improvement in efficacy between the two conjugated oligonucleotides was additionally provided by replacing part of the PS linkage with PO (ISIS 656173).

Table 50a

Factor XI protein (saline % )

Serum hepatic transaminase levels, alanine transaminase (ALT) and aspartate transaminase (AST) were measured for saline infused mice using standard protocols. Total bilirubin, total albumin, CRE and BUN were also assessed. Changes in body weight were assessed without significant changes from the saline group. ALT, AST, total bilirubin and BUN values are shown in the table below.

Table 51

Example 60: in vitro SRB targeting -1 conjugated ASO's effect

The oligonucleotides listed below were tested in a multi-dose study for antisense inhibition of SRB-1 in primary mouse hepatocytes. ISIS 353382 was included as an unconjugated standard. Each conjugate group was attached at the 3' or 5' end of each oligonucleotide by a phosphodiester linked 2'-deoxyadenosine nucleoside cleavable moiety.

Table 52

SRB Modified to target -1 ASO

Capital letters represent the nucleobase for each nucleoside and ^m C represents 5-methyl cytosine. subscript: "e" represents a 2'-MOE modified nucleoside; "d" represents β-D-2'-deoxyribonucleoside;"s" represents a phosphorothioate internucleoside linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO); And "o'" represents -OP(=O)(OH)-. Conjugate groups are in bold type.

The structure of "GalNAc ₃ -1 _a " is already shown in Example 9. The structure of “GalNAc ₃ -3a” is already shown in Example 39. The structure of “GalNAc ₃ -8a” is already shown in Example 47. The structure of “GalNAc ₃ -9a” is already shown in Example 52. The structure of the "GalNAc -6a _3" is already shown in Example 51. The structure of "GalNAc ₃ -2a" is already shown in Example 37. The structure of "GalNAc ₃ -10a" is already shown in Example 46. The structure of “GalNAc ₃ -5a” is already shown in Example 49. The structure of “GalNAc ₃ -7a” is already shown in Example 48.

cure

The oligonucleotides listed above were tested in vitro in plated primary mouse hepatocytes at a density of 25,000 cells/well and treated with 0.03, 0.08, 0.24, 0.74, 2.22, 6.67 or 20 nM modified oligonucleotides. After a treatment period of approximately 16 hours, RNA was isolated from the cells and mRNA levels were measured by quantitative real-time PCR and SRB-1 mRNA levels were adjusted for total RNA content as measured by RiboGreen®.

IC ₅₀ was calculated using standard methods and the results are presented in Table 53. These results show that under free uptake conditions in which reagents or electroporation techniques are not used to artificially promote entry of oligonucleotides into cells, oligonucleotides containing GalNAc conjugates are compared to parent oligonucleotides not containing GalNAc conjugates. (ISIS 353382) was significantly stronger in hepatocytes.

Table 53

Example 61: GalNAc ₃ Preparation of oligomeric compound 175 comprising -12

Compound 169 is commercially available. Compound 172 was prepared by adding benzyl (perfluorophenyl) glutarate to compound 171. Benzyl (perfluorophenyl) glutarate was prepared by adding PFP-TFA and DIEA in DMF to 5-(benzyloxy)-5-oxopentanoic acid. An oligomeric compound 175 comprising a GalNAc ₃ -12 conjugate group was prepared from compound 174 using the general procedure exemplified in Example 46. Konju gate group GalNAc ₃ -12 GalNAc ₃ Cluster moieties (GalNAc ₃ -12 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-. The structure of GalNAc ₃ -12 (GalNAc ₃ -12 _a -CM-) is as follows:

Example 62: GalNAc ₃ - 13 Preparation of oligomeric compound 180 comprising

Compound 176 was prepared using the general procedure shown in Example 2. An oligomeric compound 180 comprising a GalNAc ₃ -13 conjugate group was prepared from compound 177 using the general procedure exemplified in Example 49. Konju gate group GalNAc ₃ -13 GalNAc ₃ Cluster moieties (GalNAc ₃ -13 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-. The structure of GalNAc ₃ -13 (GalNAc ₃ -13 _a -CM-) is as follows:

Example 63: GalNAc ₃ Preparation of oligomeric compound 188 comprising -14

Compounds 181 and 185 are commercially available. An oligomeric compound 188 comprising a GalNAc ₃ -14 conjugate group was prepared from compound 187 using the general procedure exemplified in Example 46. Konju gate group GalNAc ₃ -14 GalNAc ₃ Cluster moieties (GalNAc ₃ -14 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-. The structure of GalNAc ₃ -14 (GalNAc ₃ -14 _a -CM-) is as follows:

Example 64: GalNAc ₃ Preparation of oligomeric compound 197 comprising -15

Compound 189 is commercially available. Compound 195 was prepared using the general procedure shown in Example 31. GalNAc ₃ -15 konju oligomeric compound 197 including the gate group, standard oligonucleotides using oligonucleotide synthesis procedure was obtained from the compound 194 and 195. Konju gate group GalNAc ₃ -15 GalNAc ₃ Cluster moieties (GalNAc ₃ -15 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-. The structure of GalNAc ₃ -15 (GalNAc ₃ -15 _a -CM-) is as follows:

Example 65: in vivo at SRB 5'- to target -1 conjugate group (GalNAc ₃ -3, 12, 13, 14, and 15 comparisons), a dose-dependent study of oligonucleotides including

The oligonucleotides listed below were tested in a dose-dependent study for antisense inhibition of SRB-1 in mice. The unconjugated ISIS 353382 is included as a standard. Each GalNAc ₃ conjugate group was attached at the 5' end of each oligonucleotide by a phosphodiester linked 2'-deoxyadenosine nucleoside cleavable moiety.

Table 54

SRB Modified to target -1 ASO

Capital letters represent the nucleobase for each nucleoside and ^m C represents 5-methyl cytosine. subscript: "e" represents a 2'-MOE modified nucleoside; "d" represents β-D-2'-deoxyribonucleoside;"s" represents a phosphorothioate internucleoside linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO); And "o'" represents -OP(=O)(OH)-. Conjugate groups are in bold type.

The structure of "GalNAc ₃ -3 _a " is already shown in Example 39. The structure of the "GalNAc ₃ -12a" is shown in Example 61. The structure of the "GalNAc ₃ -13a" is shown in Example 62. The structure of “GalNAc ₃ -14a” is shown in Example 63. The structure of “GalNAc ₃ -15a” is shown in Example 64.

cure

6-8 week old C57bl6 mice (Jackson Laboratory, Bar Harbor, ME) were injected subcutaneously once or twice with ISIS 353382, 661161, 671144, 670061, 671261, 671262, or saline at the doses shown below. Mice dosed twice received the second dose 3 days after the first dose. Each treatment group consisted of 4 animals. Mice were sacrificed 72 hours after final administration and liver SRB-1 mRNA levels were measured using real-time PCR and RiboGreen® RNA quantification reagent according to standard protocols (Molecular Probes, Inc. Eugene, OR). The results below are presented as the average percentage of SRB-1 mRNA levels for each treatment group normalized to the saline control.

As demonstrated in Table 55, treatment with antisense oligonucleotides reduced SRB-1 mRNA levels in a dose-dependent manner. No significant differences in target knockdown were observed between animals receiving single doses and animals receiving double doses (see ISIS 353382 doses 30 and 2 x 15 mg/kg; and ISIS 661161 doses 5 and 2). x 2.5 mg/kg). Antisense oligonucleotides comprising phosphodiester linked GalNAc ₃ -3, 12, 13, 14, and 15 conjugates showed substantial improvement in efficacy compared to unconjugated antisense oligonucleotides (ISIS 335382).

Table 55

SRB -One mRNA ( % brine)

Serum hepatic transaminase levels, alanine transaminase (ALT) and aspartate transaminase (AST) were measured for saline infused mice using standard protocols. Total bilirubin and BUN were also assessed. Changes in body weight were assessed without significant differences from the saline group (data not shown). ALT, AST, total bilirubin and BUN values are shown in Table 56 below.

Table 56

Example 66: 5' - GalNAc ₃ containing the cluster SRB by oligonucleotides targeting -1 in vivo antisense Various cutting possibilities for restraint moiety effect

The oligonucleotides listed below were tested in a dose-dependent study for antisense inhibition of SRB-1 in mice. Each GalNAc ₃ conjugate group was attached at the 5' end of each oligonucleotide by a phosphodiester linked nucleoside cleavable moiety (CM).

Table 57

SRB Modified to target -1 ASO

Capital letters represent the nucleobase for each nucleoside and ^m C represents 5-methyl cytosine. subscript: "e" represents a 2'-MOE modified nucleoside; "d" represents β-D-2'-deoxyribonucleoside;"s" represents a phosphorothioate internucleoside linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO); And "o'" represents -OP(=O)(OH)-. Conjugate groups are in bold type.

The structure of "GalNAc ₃ -3 _a " is already shown in Example 39. The structure of the "GalNAc ₃ -13a" is shown in Example 62.

cure

6-8 week old C57bl6 mice (Jackson Laboratory, Bar Harbor, ME) were injected once subcutaneously with ISIS 661161, 670699, 670700, 670701, 671165, or saline at the doses shown below. Each treatment group consisted of 4 animals. Mice were sacrificed 72 hours after final administration and liver SRB-1 mRNA levels were measured using real-time PCR and RiboGreen® RNA quantification reagent according to standard protocols (Molecular Probes, Inc. Eugene, OR). The results below are presented as the average percentage of SRB-1 mRNA levels for each treatment group normalized to the saline control.

As demonstrated in Table 58, treatment with antisense oligonucleotides reduced SRB-1 mRNA levels in a dose-dependent manner. Antisense oligonucleotides containing all of the various cleavable moieties showed similar efficacy.

Table 58

SRB -One mRNA ( % brine)

Serum hepatic transaminase levels, alanine transaminase (ALT) and aspartate transaminase (AST) were measured for saline infused mice using standard protocols. Total bilirubin and BUN were also assessed. Changes in body weight were assessed without significant differences from the saline group (data not shown). ALT, AST, total bilirubin and BUN values are shown in Table 56 below.

Table 59

Example 67: GalNAc ₃ - 16 Preparation of oligomeric compound 199 comprising

The GalNAc ₃ -16 konju oligomeric compound 199 including the gate group was prepared using the general procedure illustrated in Examples 7 and 9. Konju gate group GalNAc ₃ -16 GalNAc ₃ Cluster moieties (GalNAc ₃ -16 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-. The structure of GalNAc ₃ -16 (GalNAc ₃ -16 _a -CM-) is shown below:

Example 68: GalNAc ₃ - 17 Preparation of oligomeric compound 200 comprising

An oligomeric compound 200 containing a GalNAc ₃ -17 conjugate group was prepared using the general procedure exemplified in Example 46. The GalNAc ₃ cluster portion of the conjugate group GalNAc _{3 -17 (GalNAc 3} -17 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-. The structure of GalNAc ₃ -17 (GalNAc ₃ -17 _a -CM-) is as follows:

Example 69: GalNAc ₃ - 18 Preparation of oligomeric compound 201 comprising

Oligomeric Compound 201 comprising a GalNAc ₃ -18 conjugate group was prepared using the general procedure exemplified in Example 46. The GalNAc ₃ cluster portion of the conjugate group GalNAc _{3 -18 (GalNAc 3} -18 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-. The structure of GalNAc ₃ -18 (GalNAc ₃ -18 _a -CM-) is as follows:

Example 70: GalNAc ₃ Preparation of oligomeric compound 204 comprising -19

GalNAc ₃ -19 konju using a general procedure illustrated oligomeric compound 204 including the gate group in Example 52 was prepared from compound 64. Konju gate group GalNAc ₃ -19 GalNAc ₃ Cluster moieties (GalNAc ₃ -19 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-. The structure of GalNAc ₃ -19 (GalNAc ₃ -19 _a -CM-) is as follows:

Example 71: GalNAc ₃ - 20 Preparation of oligomeric compound 210 comprising

Compound 205 was prepared by adding PFP-TFA and DIEA to 6-(2,2,2-trifluoroacetamido)hexanoic acid in acetonitrile prepared by adding triflic anhydride to 6-aminohexanoic acid did. The reaction mixture ^{was heated to 80 °} C, then cooled to room temperature. GalNAc ₃ -20 konju to the oligomeric compound 210 including the gate groups using the general procedure exemplified in Example 52 was prepared from compound 208. Konju gate group GalNAc ₃ -20 GalNAc ₃ Cluster portion (GalNAc ₃ -20 _a) is combined with any cuttable moiety can provide a wide range of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-. The structure of _{GalNAc 3 -20 (GalNAc 3 -20 a} -CM-) is as follows:

Example 72: GalNAc ₃ - 21 Preparation of oligomeric compound 215 comprising

Compound 211 is commercially available. An oligomeric compound 215 containing a GalNAc ₃ -21 conjugate group was prepared from compound 213 using the general procedure exemplified in Example 52. Konju gate group GalNAc ₃ -21 GalNAc ₃ The cluster moiety (GalNAc ₃ -21 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-. The structure of GalNAc ₃ -21 (GalNAc ₃ -21 _a -CM-) is as follows:

Example 73: GalNAc ₃ Preparation of oligomeric compound 221 comprising -22

Compound 220 was prepared from compound 219 using diisopropylammonium tetrazolide. An oligomeric compound 221 containing a GalNAc ₃ -21 conjugate group was prepared from compound 220 using the general procedure exemplified in Example 52. Konju gate group GalNAc ₃ -22 GalNAc ₃ The cluster moiety (GalNAc ₃ -22 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. In certain embodiments, the cleavable moiety is -P(=O)(OH)-A _d -P(=O)(OH)-. The structure of GalNAc ₃ -22 (GalNAc ₃ -22 _a -CM-) is as follows:

Example 74: 5' - GalNAc ₃ the conjugate containing SRB by oligonucleotides targeting -1 in vivo antisense Various cutting possibilities for restraint moiety effect

The oligonucleotides listed below were tested in a dose-dependent study for antisense inhibition of SRB-1 in mice. Each GalNAc ₃ conjugate group was attached at the 5' end of each oligonucleotide.

Table 60

SRB Modified to target -1 ASO

In all tables, uppercase letters represent the nucleobase for each nucleoside and ^m C represents 5-methyl cytosine. subscript: "e" represents a 2'-MOE modified nucleoside; "d" represents β-D-2'-deoxyribonucleoside;"s" represents a phosphorothioate internucleoside linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO); And "o'" represents -OP(=O)(OH)-. Conjugate groups are in bold type.

The structure of "GalNAc ₃ -3 _a " is already shown in Example 39. The structure of GalNAc ₃ -17a is already shown in Example 68, and _{the structure of GalNAc 3} -18a is shown in Example 69.

cure

6-8 week old C57BL/6 mice (Jackson Laboratory, Bar Harbor, ME) were injected once subcutaneously with the oligonucleotides listed in Table 60 or saline at the doses shown below. Each treatment group consisted of 4 animals. Mice were sacrificed 72 hours after final dosing and SRB-1 mRNA levels were measured using real-time PCR and RiboGreen® RNA quantification reagent according to standard protocols (Molecular Probes, Inc. Eugene, OR). The results below are presented as the average percentage of SRB-1 mRNA levels for each treatment group normalized to the saline control.

As demonstrated in Table 61, treatment with antisense oligonucleotides reduced SRB-1 mRNA levels in a dose-dependent manner. Antisense oligonucleotides with GalNAc conjugates showed similar potency and were significantly stronger than the parental oligonucleotides without GalNAc conjugates.

Table 61

SRB -One mRNA ( % brine)

Serum hepatic transaminase levels, alanine transaminase (ALT) and aspartate transaminase (AST) were measured for saline infused mice using standard protocols. Total bilirubin and BUN were also assessed. Changes in body weight were assessed without significant changes from the saline group (data not shown). ALT, AST, total bilirubin and BUN values are shown in Table 62 below.

Table 62

Example 75: 5 '- conjugate of an oligonucleotide comprising a group pharmacodynamic analyze

The PK of ASO in Tables 54, 57, and 60 above was evaluated using liver samples obtained according to the treatment procedures described in Examples 65, 66, and 74. Liver samples were minced, extracted using standard protocols and analyzed by IP-HPLC-MS with internal standards. Combined tissue levels (μg/g) of all metabolites were determined by incorporating the appropriate UV peaks (“parent” in this case Isis number 353382) and tissue levels of full-length ASO missing the conjugate were compared with the appropriate extracted ion chromatography. It was measured using grams (EIC).

Table 63

Hepatic PK analysis

The results of Table 63 above, especially considering the difference in administration between the oligonucleotides depending on the presence or absence of the _{GalNAc 3} conjugate group, 72 hours after administration of the oligonucleotide, the parent oligo that does not contain the _{GalNAc 3 conjugate group (ISIS 353382).} It shows that the liver tissue level of oligonucleotides containing _{GalNAc 3 conjugate groups is greater than that of nucleotides.} Moreover, by 72 h, _{40-98% of each oligonucleotide containing the GalNAc 3} conjugate group was metabolized to yield the parent compound, _{indicating that the GalNAc 3} conjugate group was cleaved from the oligonucleotide.

Example 76: GalNAc ₃ - 23 Preparation of oligomeric compound 230 comprising

Compound 222 is commercially available. 44.48 ml (0.33 mol) of compound 222 was treated with tosyl chloride (25.39 g, 0.13 mol) in pyridine (500 mL) for 16 h. The reaction was then evaporated to an oil, dissolved in EtOAc and washed with water, saturated NaHCO ₃ , brine and dried over _{Na 2} SO _{4 .} Ethyl acetate was concentrated to dryness and purified by column chromatography, eluting with EtOAc/hexanes (1:1) then _{10% methanol in CH 2} Cl ₂ to give compound 223 as a colorless oil. LCMS and NMR were consistent with the structure. 10 g (32.86 mmol) of 1-tosyltriethylene glycol (Compound 223) was treated with sodium azide (10.68 g, 164.28 mmol) in DMSO (100 mL) at room temperature for 17 h. Then the reaction mixture was poured into water and extracted with EtOAc. The organic layer was washed 3 times with water and dried _{over Na 2} SO _{4 .} The organic layer was concentrated to dryness to obtain 5.3 g of compound 224 (92%). LCMS and NMR were consistent with the structure. 1-azidotriethylene glycol (compound 224, 5.53 g, 23.69 mmol) and compound 4 (6 g, 18.22 mmol) were mixed with 4A molecular sieve (5 g), and TMSOTf (1.65 ml, 9.11 mmol) in dichloromethane (100 mL) treated under an inert atmosphere. After 14 h, the reaction was filtered off and the organic layer was _{washed with saturated NaHCO 3} , brine and dried over _{Na 2} SO _{4 .} The organic layer was concentrated to dryness, purified by column chromatography, and eluted with a gradient of 2-4% methanol in dichloromethane to give compound 225. LCMS and NMR were consistent with the structure. Compound 225 (11.9 g, 23.59 mmol) was hydrogenated over Perlman catalyst in EtOAc/methanol (4:1, 250 mL). After 8 hours, the catalyst was removed by filtration, the solvent was removed and dried to give compound 226. LCMS and NMR were consistent with the structure.

To yield compound 227, a solution of nitromethanetrispropionic acid (4.17 g, 15.04 mmol) and Hunig's base (10.3 ml, 60.17 mmol) in DMF (100 mL) was dissolved in pentafluorotrifluoro acetate (9.05 ml, 52.65 mmol) dealt with dropwise. After 30 min, the reaction was poured into ice water and extracted with EtOAc. The organic layer was washed with water, brine and dried _{over Na 2} SO _{4 .} The organic layer was concentrated to dryness and then recrystallized from heptane to give compound 227 as a white solid. LCMS and NMR were consistent with the structure. Compound 227 (1.5 g, 1.93 mmol) and compound 226 (3.7 g, 7.74 mmol) were stirred in acetonitrile (15 mL) at room temperature for 2 h. The reaction was then evaporated to dryness and purified by column chromatography eluting with a gradient of 2-10% methanol in dichloromethane to give compound 228. LCMS and NMR were consistent with the structure. Compound 228 (1.7 g, 1.02 mmol) was treated with Raney Nickel (ca. 2 g wet) in ethanol (100 mL) in an atmosphere of hydrogen. After 12 hours, the catalyst was removed by filtration and the organic layer was evaporated to give a solid, which was used directly in the next step. LCMS and NMR were consistent with the structure. This solid (0.87 g, 0.53 mmol) was treated with benzylglutaric acid (0.18 g, 0.8 mmol), HBTU (0.3 g, 0.8 mmol) and DIEA (273.7 μl, 1.6 mmol) in DMF (5 mL). After 16 h, DMF was removed at 65 under reduced pressure to give an oil, which was dissolved in dichloromethane. The organic layer was _{washed with saturated NaHCO 3} , brine and dried over _{Na 2} SO _{4 .} After evaporation of the organic layer, the compound was purified by column chromatography and eluted with a gradient of 2-20% methanol in dichloromethane to give the coupled product. LCMS and NMR were consistent with the structure. The benzyl ester was deprotected with a Perlman catalyst under a hydrogen atmosphere for 1 h. Then the catalyst was removed by filtration, the solvent was removed and dried to obtain an acid. LCMS and NMR were consistent with the structure. The acid (486 mg, 0.27 mmol) was dissolved in dry DMF (3 mL). Pyridine (53.61 μl, 0.66 mmol) was added and the reaction was purged with argon. Pentafluorotrifluoro acetate (46.39 μl, 0.4 mmol) was added slowly to the reaction mixture. The color of the reaction changed from pale yellow to dark red, and a light smoke was formed, which was removed with a stream of argon. The reaction was allowed to stir at room temperature for 1 h (reaction completion was confirmed by LCMS). The solvent was removed at 70° C. under reduced pressure (rotovap). The residue was diluted with DCM and washed with 1N NaHSO ₄ , brine, and washed again with saturated sodium bicarbonate and brine. The organics were _{dried over Na 2} SO ₄ , filtered and concentrated to dryness to give 225 mg of compound 229 as a brittle yellow foam. LCMS and NMR were consistent with the structure.

Oligomeric compound 230 comprising a GalNAc ₃ -23 conjugate group was prepared from compound 229 using the general procedure demonstrated in Example 46. GalNAc ₃ -23 konju gate group ₃ GalNAc The cluster moiety (GalNAc ₃ -23 _a ) can be combined with any cleavable moiety to provide a variety of conjugate groups. The structure of GalNAc ₃ -23 (GalNAc ₃ -23 _a -CM) is as follows:

Example 77: GalNAc ₃ a conjugate containing SRB by oligonucleotides targeting -1 in vivo antisense control

The oligonucleotides listed below were tested in a dose-dependent study for antisense inhibition of SRB-1 in mice.

Table 64

SRB Modified to target -1 ASO

The structure of GalNAc3-1a is already shown in Example 9, the structure of GalNAc3-3a is shown in Example 39, the structure of GalNAc3-9a is shown in Example 52, and the structure of GalNAc3-10a is shown in Example 46 The structure of GalNAc3-19a is shown in Example 70, the structure of GalNAc3-20a is shown in Example 71, and the structure of GalNAc3-23a is shown in Example 76.

cure

Each of 6-8 week old C57BL/6 mice (Jackson Laboratory, Bar Harbor, ME) was injected once subcutaneously with the oligonucleotides listed in Table 64 or saline at the doses shown below. Each treatment group consisted of 4 animals. Mice were sacrificed 72 hours after final dosing and SRB-1 mRNA levels were measured using real-time PCR and RiboGreen® RNA quantification reagent according to standard protocols (Molecular Probes, Inc. Eugene, OR). The results below are presented as the average percentage of SRB-1 mRNA levels for each treatment group normalized to the saline control.

As demonstrated in Table 65, treatment with antisense oligonucleotides reduced SRB-1 mRNA levels in a dose-dependent manner.

Table 65

SRB -One mRNA ( % brine)

Serum hepatic transaminase levels, alanine transaminase (ALT) and aspartate transaminase (AST) were also measured using standard protocols. Total bilirubin and BUN were also assessed. Changes in body weight were assessed and there were no significant changes from the saline group (data not shown). ALT, AST, total bilirubin and BUN values are shown in Table 66 below.

Table 66

Example 78: GalNAc ₃ a conjugate containing angiotensinogen by targeting oligonucleotides in vivo antisense control

The oligonucleotides listed below were tested in a dose-dependent study for antisense inhibition of angiotensinogen (AGT) in normotensive Sprague Dowry rats.

Table 67

AGT modified to target ASO

The structure of "GalNAc ₃ -1 _a " is already shown in Example 9.

cure

Each 6-week-old, male Sprague Dowry rat was injected subcutaneously once per week with the oligonucleotides listed in Table 67 or PBS at the doses shown below for a total of 3 doses. Each treatment group consisted of 4 animals. Rats were sacrificed 72 hours after the last dose. AGT liver mRNA levels were measured using real-time PCR and RiboGreen® RNA Quantification Reagent (Molecular Probes, Inc. Eugene, OR) according to standard protocols. AGT plasma protein levels were measured using total angiotensinogen ELISA (catalog # JP27412, IBL International, Toronto, ON) with 1:20,000 diluted plasma. The results below are presented as mean percent of AGT mRNA levels in liver or plasma of AGT protein levels in each treatment group normalized to PBS control.

As demonstrated in Table 68, treatment with antisense oligonucleotides reduced AGT liver mRNA and plasma protein levels in a dose-dependent manner, and oligonucleotides comprising GalNAc conjugates were more effective than parent oligonucleotides without GalNAc conjugates. significantly stronger.

Table 68

AGT liver mRNA and plasma protein levels

Hepatic transaminase levels, alanine transaminase (ALT) and aspartate transaminase (AST) in plasma and body weight were also measured at sacrifice using standard protocols. The results are shown in Table 69 below.

Table 69

liver transaminase levels and rat weight

Example 79: GalNAc ₃ a conjugate containing APOC of oligonucleotides targeting -III in vivo duration of action

The oligonucleotides listed in Table 70 below were tested in a single dose study for duration of action in mice.

Table 70

APOC Modified to target -III ASO

The structure of GalNAc ₃ -1 _a is already shown in Example 9, the structure of GalNAc ₃ -3 _a is shown in Example 39, _{the structure of GalNAc 3-7 a} is shown in Example 48, and GalNAc ₃ -10 the structure of _a may appear in example 46, the structure of GalNAc ₃ -13 _a are set forth in example 62.

cure

Each of 6-8 week old transgenic mice expressing human APOC-III were injected subcutaneously once with the oligonucleotides listed in Table 70 or PBS. Each treatment group consisted of 3 animals. Baseline was determined by drawing blood before dosing and at 72 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, and 6 weeks after dosing. Plasma triglyceride and APOC-III protein levels were measured as described in Example 20. The results below are presented as the mean percent of plasma triglyceride and APOC-III levels for each treatment group normalized to baseline levels, indicating that oligonucleotides comprising a group of GalNAc conjugates have a simulated dose comprising a group of GalNAc conjugates. It shows that even at 3 times the dose of the oligonucleotide, it exhibits a longer duration of action than the parent oligonucleotide without the conjugate group (ISIS 304801).

Table 71

Plasma in transgenic mice triglycerides and APOC -III protein level

Example 80: GalNAc ₃ a conjugate Alpha-1 containing Antitrypsin (A1AT) by targeting oligonucleotides in vivo antisense control

The oligonucleotides listed in Table 72 below were tested in a study for dose-dependent inhibition of A1AT in mice.

Table 72

A1AT modified to target ASO

The structure of GalNAc ₃ -1 _a is already shown in Example 9, the structure of GalNAc ₃ -3 _a is shown in Example 39, _{the structure of GalNAc 3-7 a} is shown in Example 48, and GalNAc ₃ -10 the structure of _a may appear in example 46, and the structure of GalNAc ₃ -13 _a are set forth in example 62.

cure

Each 6-week-old, male C57BL/6 mouse (Jackson Laboratory, Bar Harbor, ME) was injected subcutaneously once per week with the oligonucleotides listed in Table 72 or PBS at the doses shown below, for a total of 3 doses. . Each treatment group consisted of 4 animals. Mice were sacrificed 72 hours after the last dose. A1AT liver mRNA levels were measured using real-time PCR and RiboGreen® RNA Quantification Reagent (Molecular Probes, Inc. Eugene, OR) according to standard protocols. A1AT plasma protein levels were determined using mouse alpha 1-antitrypsin ELISA (catalog #41-A1AMS-E01, Alpco, Salem, NH). The results below are presented from the mean percent of A1AT liver mRNA and plasma protein levels for each treatment group normalized to the PBS control.

As demonstrated in Table 73, treatment with antisense oligonucleotides reduced A1AT liver mRNA and A1AT plasma protein levels in a dose-dependent manner. Oligonucleotides containing the GalNAc conjugate were significantly stronger than the parent (ISIS 476366).

Table 73

A1AT liver mRNA and plasma protein levels

Hepatic transaminase and BUN levels in plasma were measured at sacrifice using standard protocols. Body weight and organ weight were also measured. The results are shown in Table 74 below. Body weight is shown as a % of baseline. Organ weights are shown as % of body weight relative to the PBS control group.

Table 74

Example 81: GalNAc ₃ containing the cluster A1AT of the targeted oligonucleotide in vivo duration of action

The oligonucleotides listed in Table 72 were tested in a single dose study for duration of action in mice.

cure

Each 6-week-old, male C57BL/6 mouse was injected subcutaneously once with the oligonucleotides listed in Table 72 or PBS. Each treatment group consisted of 4 animals. Blood was drawn the day before dosing to determine baseline and at 5, 12, 19, and 25 days post-dose. Plasma A1AT protein levels were determined via ELISA (Reference Example 80). Results below are presented as mean percent of plasma A1AT protein levels for each treatment group normalized to baseline levels. These results show that the oligonucleotides containing the GalNAc conjugate were stronger and had a longer duration of action than the parent without the GalNAc conjugate (ISIS 476366). Moreover, oligonucleotides comprising a 5'-GalNAc conjugate (ISIS 678381, 678382, 678383, and 678384) have a longer duration of action than oligonucleotides comprising a 3'-GalNAc conjugate (ISIS 656326). In general, the stronger the stronger.

Table 75

plasma in mice A1AT protein level

Example 82: GalNAc ₃ a conjugate containing SRB targeting -1 oligogne by cleotide in vitro antisense control

Primary mouse liver hepatocytes were seeded 2 h prior to treatment at 15,000 cells/well in 96 well plates. Oligonucleotides listed in Table 76 were added in Williams E medium at 2, 10, 50, or 250 nM and cells were incubated overnight at 37 ^o C in 5% CO ₂ . Cells were lysed for 16 h following oligonucleotide addition, and total RNA was purified using an RNease 3000 BioRobot (Qiagen). SRB-1 mRNA levels were measured using real-time PCR and RiboGreen® RNA Quantification Reagent (Molecular Probes, Inc. Eugene, OR) according to standard protocols. IC ₅₀ values were measured using Prism 4 software (GraphPad). These results show that oligonucleotides containing various different GalNAc conjugate groups and various different cleavable moieties are significantly stronger in in vitro free uptake experiments than parent oligonucleotides without GalNAc conjugate groups (ISIS 353382 and 666841). show that

Table 76

in vitro SRB -1 inhibition of expression

The structure of GalNAc ₃ -1 _a is already shown in Example 9, the structure of GalNAc ₃ -3 _a is shown in Example 39, _{the structure of GalNAc 3} -5 _a is shown in Example 49, and GalNAc ₃ -6 the structure of _a may appear in example 51, the structure of _a GalNAc ₃ -7 are shown and in example 48, the structure of _a GalNAc ₃ -8 has shown in example 47, the structure of GalNAc are carried out _{a 3} -9 and shown in example 52, the structure of GalNAc ₃ -10 _a are measured and shown in example 46, the structure of _a GalNAc -12 ₃ has shown in example 61, the structure of _a GalNAc ₃ -13 may appear in example 62 , _{The structure of GalNAc 3} -14 _a is shown in Example 63, the structure of GalNAc ₃ -15 _a is shown in Example 64, _{the structure of GalNAc 3} -17 _a is shown in Example 68, and GalNAc ₃ -18 a the structure of _a may appear in the example 69, the structure of _a GalNAc -19 ₃ has shown in example 70, the structure of _a GalNAc ₃ -20 may appear in example 71, the structure of GalNAc are carried out _{a 3} -23 It is shown in Example 76.

Example 83: GalNAc ₃ by oligonucleotides targeting factor XI comprising clusters in vivo antisense control

The oligonucleotides listed in Table 77 below were tested in a study for dose-dependent inhibition of factor XI in mice.

Table 77

Modified oligonucleotides targeting factor XI

The structure of GalNAc ₃ -1 _a is already shown in Example 9, the structure of GalNAc ₃ -3 _a is shown in Example 39, _{the structure of GalNAc 3-7 a} is shown in Example 48, and GalNAc ₃ -10 the structure of _a may appear in example 46, and the structure of GalNAc ₃ -13 _a are set forth in example 62.

cure

Each of 6-8 week old mice was injected subcutaneously once per week with the oligonucleotides listed below or PBS at the doses shown below, for a total of 3 doses. Each treatment group consisted of 4 animals. Mice were sacrificed 72 hours after the last dose. Factor XI liver mRNA levels were measured using real-time PCR and normalized to cyclophilin according to standard protocols. Liver transaminase, BUN, and bilirubin were also measured. Results below are presented as average percentages for each treatment group normalized to PBS control.

As demonstrated in Table 78, treatment with antisense oligonucleotides reduced Factor XI liver mRNA in a dose-dependent manner. These results show that the oligonucleotides containing the GalNAc conjugate were stronger than the parent without the GalNAc conjugate (ISIS 404071). Moreover, oligonucleotides containing 5'-GalNAc conjugates (ISIS 663086, 678347, 678348, and 678349) were even stronger than oligonucleotides containing 3'-GalNAc conjugates (ISIS 656173).

Table 78

Factor XI liver mRNA , liver transaminase, BUN, and bilirubin levels

Example 84: GalNAc ₃ a conjugate of oligonucleotides targeting factor XI, including in vivo duration of action

The oligonucleotides listed in Table 77 were tested in a single dose study for duration of action in mice.

cure

Each of the 6-8 week old mice was injected subcutaneously once with the oligonucleotides listed in Table 77 or PBS. Each treatment group consisted of 4 animals. Blood was drawn by tail bleed the day before dosing to determine baseline and measured at 3, 10, and 17 days post-dose. Plasma Factor XI protein levels were measured with Factor XI capture and biotinylated detection antibodies (R&D SYSTEMS, Minneapolis, MN (catalogs #AF2460 and #BAF2460, respectively)) and OptEIA reagent set B (catalog #550534, BD Biosciences, San Jose, CA) was measured by ELISA. Results below are presented as mean percent of plasma Factor XI protein levels for each treatment group normalized to baseline levels. These results show that the oligonucleotides containing the GalNAc conjugate were stronger with a longer duration of action than the parent without the GalNAc conjugate (ISIS 404071). Moreover, oligonucleotides comprising a 5'-GalNAc conjugate (ISIS 663086, 678347, 678348, and 678349) with a longer duration of action than oligonucleotides comprising a 3'-GalNAc conjugate (ISIS 656173) stronger and stronger.

Table 79

Plasma Factor XI Protein Levels in Mice

Example 85: GalNAc ₃ a conjugate containing SRB by oligonucleotides targeting -1 in vivo antisense control

The oligonucleotides listed in Table 76 were tested in a dose-dependent study for antisense inhibition of SRB-1 in mice.

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Each of 6-8 week old C57BL/6 mice was injected subcutaneously once per week at the doses shown below, for a total of 3 doses, with the oligonucleotides listed in Table 76 or saline. Each treatment group consisted of 4 animals. Mice were sacrificed 48 h after final dosing and SRB-1 mRNA levels were measured using real-time PCR and RiboGreen® RNA quantification reagent according to standard protocols (Molecular Probes, Inc. Eugene, OR). Results below are presented as mean percent of liver SRB-1 mRNA levels for each treatment group normalized to saline control.

Table 80 ?? 81, treatment with antisense oligonucleotides reduced SRB-1 mRNA levels in a dose-dependent manner.

Table 80

in the middle SRB -One mRNA

Table 81

in the middle SRB -One mRNA

Liver transaminase levels, total bilirubin, BUN, and body weight were also measured using standard protocols. The average values for each treatment group are shown in Table 82 below.

Table 82

Example 86: GalNAc ₃ containing the cluster TTR by targeting oligonucleotides in vivo antisense control

The oligonucleotides listed in Table 83 below were tested in a dose-dependent study for antisense inhibition of human transthyretin (TTR) in transgenic mice expressing the human TTR gene.

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Each of the 8-week-old TTR transgenic mice was injected subcutaneously once per week for 3 weeks, for a total of 3 doses, with the oligonucleotides and doses listed in the table below or PBS. Each treatment group consisted of 4 animals. Mice were sacrificed 72 hours after the last dose. Tail bleeds were performed at various time points throughout the experiment, and plasma TTR protein, ALT, and AST levels were measured and reported in Tables 85-87. After the animals were sacrificed, plasma ALT, AST, and human TTR levels were measured as in body weight, organ weight, and liver human TTR mRNA levels. TTR protein levels were measured using a clinical analyzer (AU480, Beckman Coulter, CA). Real-time PCR and RiboGreen® RNA quantification reagent were used according to standard protocols to measure liver human TTR mRNA levels (Molecular Probes, Inc. Eugene, OR). The results presented in Tables 84-87 are the mean values for each treatment group. mRNA levels are the mean values for the mean of the PBS group. Plasma protein levels are the mean values for the mean values of the PBS group at baseline. Body weight is the mean percent change in body weight from baseline until sacrifice for each individual treatment group. The organ weights shown are normalized to the animal's body weight, and then the average normalized organ weight of each treatment group is plotted against the average normalized organ weight of the PBS group.

In Tables 84-87, "BL" represents the baseline, measurement taken immediately before the first dose. Table 84 ?? 85, treatment with antisense oligonucleotides reduced TTR expression levels in a dose-dependent manner. Oligonucleotides containing the GalNAc conjugate were stronger than the parent without the GalNAc conjugate (ISIS 420915). Moreover, GalNAc conjugates and oligonucleotides comprising mixed PS/PO internucleoside linkages are GalNAc conjugates and oligonucleotides comprising complete PS linkages.

Table 83

human TTR Targeted oligonucleotides

A legend to Table 85 can be found in Example 74. The structure of “GalNAc ₃ -1” is shown in Example 9. The structure of “GalNAc ₃ -3 _a " is shown in Example 39. The structure of "GalNAc ₃ -7 _a " is shown in Example 48. The structure of “GalNAc ₃ -10 _a " is shown in Example 46. The structure of "GalNAc ₃ -13 _a " is shown in Example 62. The structure of “GalNAc ₃ -19 _a ” is shown in Example 70.

Table 84

in vivo human TTR's antisense control

Table 85

in vivo human TTR's antisense control

Table 86

transaminase Levels, Weight Changes, and Organ Weights

Table 87

transaminase Levels, Weight Changes, and Organ Weights

Example 87: GalNAc ₃ containing the cluster TTR by a single dose of the targeted oligonucleotide in vivo duration of action

ISIS numbers 420915 and 660261 (see Table 83) were tested in a single dose study for duration of action in mice. ISIS numbers 420915, 682883, and 682885 (see Table 83) were also tested in a single dose study for duration of action in mice.

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Each of 8-week-old, male transgenic mice expressing human TTR were injected subcutaneously once with 100 mg/kg ISIS No. 420915 or 13.5 mg/kg ISIS No. 660261. Each treatment group consisted of 4 animals. Tail bleeds were performed prior to dosing to determine baseline and measured 3, 7, 10, 17, 24, and 39 days post dosing. Plasma TTR protein levels were measured as described in Example 86. Results below are presented as mean percent of plasma TTR levels for each treatment group normalized to baseline levels.

Table 88

plasma TTR protein level

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Each of the female transgenic mice expressing human TTR was injected once subcutaneously with 100 mg/kg ISIS No. 420915, 10.0 mg/kg ISIS No. 682883, or 10.0 mg/kg 682885. Each treatment group consisted of 4 animals. Tail bleeds were performed prior to dosing to determine baseline and measured 3, 7, 10, 17, 24, and 39 days post dosing. Plasma TTR protein levels were measured as described in Example 86. Results below are presented as mean percent of plasma TTR levels for each treatment group normalized to baseline levels.

Table 89

plasma TTR protein level

The results in Tables 88 and 89 show that the oligonucleotide comprising the GalNAc conjugate is stronger with a longer duration than the parent oligonucleotide without the conjugate (ISIS 420915).

Example 88: GalNAc ₃ a conjugate containing smn by targeting oligonucleotides in vivo splicing control

The oligonucleotides listed in Table 90 were tested for splicing regulation of human survival of motor neurons (SMNs) in mice.

Table 90

smn modified to target ASO

The structure of "GalNAc ₃ -7 _a " is already shown in Example 48. "X" represents the 5' primary amine calculated by Gene Tools (Philomath, OR), and GalNAc _{3-7 b is GalNAc 3-7 a} without the -NH-C6-O moiety of the linker as shown below. Represents the structure:

.

.

ISIS numbers 703421 and 703422 are morpholino oligonucleotides, wherein each nucleotide of the two oligonucleotides is a morpholino nucleotide.

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Six-week-old transgenic mice expressing human SMN were injected once subcutaneously with the oligonucleotides listed in Table 91 or saline. Each treatment group consisted of 2 males and 2 females. Mice were sacrificed 3 days after administration and liver human SMN mRNA levels with and without exon 7 were measured using real-time PCR according to standard protocols. Total RNA was measured using RiboGreen reagent. SMN mRNA levels were normalized to total mRNA and further normalized to the mean of the saline treatment group. The resulting average ratios of SMN mRNA with exon 7 to SMN mRNA missing exon 7 are shown in Table 91. These results show that a fully modified oligonucleotide that modulates splicing and contains a GalNAc conjugate is significantly stronger in altering splicing in the liver than the parent oligonucleotide without a GlaNAc conjugate. Moreover, this trend holds for multiple modification chemistries involving 2′-MOE and morpholino modified oligonucleotides.

Table 91

in vivo at human smn Effect of targeting oligonucleotides

Example 89: GalNAc ₃ a conjugate containing apolipoprotein A ( spore (a)) by an oligonucleotide targeting in vivo antisense control

The oligonucleotides listed in Table 92 below were tested in a study for dose-dependent inhibition of apo(a) in transgenic mice.

Table 92

apo (a) modified to target ASO

The structure of “GalNAc ₃ -7 _a ” is shown in Example 48.

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Eight-week-old, female C57BL/6 mice (Jackson Laboratory, Bar Harbor, ME) were each injected subcutaneously once per week with the oligonucleotides listed in Table 92 or PBS at the doses shown below, for a total of 6 doses. . Each treatment group consisted of 3-4 animals. Tail bleeds were performed the day before the first dose and weekly following each dose to measure plasma apo(a) protein levels. Mice were sacrificed 2 days after the last administration. Apo(a) liver mRNA levels were measured using real-time PCR and RiboGreen® RNA Quantification Reagent (Molecular Probes, Inc. Eugene, OR) according to standard protocols. Apo(a) plasma protein levels were measured using ELISA and liver transaminase levels were measured. The mRNA and plasma protein results in Table 93 are presented as treatment group mean percent versus PBS treated group. Plasma protein levels were further normalized to the baseline (BL) values of the PBS group. Mean absolute transaminase levels and body weights (% relative to baseline mean) are reported in Table 94.

As demonstrated in Table 93, treatment with oligonucleotides reduced apo(a) liver mRNA and plasma protein levels in a dose-dependent manner. Moreover, the oligonucleotides comprising the GalNAc conjugate were significantly stronger with a longer duration of action than the parent oligonucleotides without the GalNAc conjugate. As demonstrated in Table 94, transaminase levels and body weight were not affected by the oligonucleotides, indicating that the oligonucleotides were well tolerated.

Table 93

spore (a) liver mRNA and plasma protein levels

Table 94

Example 90: GalNAc ₃ containing the cluster TTR by targeting oligonucleotides in vivo antisense control

The oligonucleotides listed in Table 95 below were tested in a dose-dependent study for antisense inhibition of human transthyretin (TTR) in transgenic mice expressing the human TTR gene.

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Each of the TTR transgenic mice was injected subcutaneously once per week for 3 weeks, for a total of 3 doses, with the oligonucleotides and doses listed in Table 96 or PBS. Each treatment group consisted of 4 animals. Prior to the first dose, a tail bleed was performed to measure plasma TTR protein levels at baseline (BL). Mice were sacrificed 72 hours after the last dose. TTR protein levels were measured using a clinical analyzer (AU480, Beckman Coulter, CA). Real-time PCR and RiboGreen® RNA quantification reagent were used according to standard protocols to measure liver human TTR mRNA levels (Molecular Probes, Inc. Eugene, OR). The results presented in Table 96 are the mean values for each treatment group. mRNA levels are the mean values for the mean of the PBS group. Plasma protein levels are the mean values of the mean values of the PBS group at baseline. “BL” refers to the baseline, measurement taken immediately prior to the first dose. As demonstrated in Table 96, treatment with antisense oligonucleotides reduced TTR expression levels in a dose-dependent manner. Oligonucleotides containing a GalNAc conjugate were stronger than the parent without the GalNAc conjugate (ISIS 420915), and oligonucleotides containing a phosphodiester or deoxyadenosine cleavable moiety were more potent than the parent without the conjugate. showed significant improvement (see reference ISIS numbers 682883 and 666943 vs 420915 and examples 86 and 87).

Table 95

human TTR Targeted oligonucleotides

A legend to Table 95 can be found in Example 74. The structure of “GalNAc ₃ -3 _a " is shown in Example 39. The structure of "GalNAc ₃ -7 _a " is shown in Example 48. The structure of “GalNAc ₃ -10 _a " is shown in Example 46. The structure of "GalNAc ₃ -13 _a " is shown in Example 62.

Table 96

in vivo human TTR's antisense control

Example 91: in non-human primates GalNAc3 a conjugate by oligonucleotides targeting factor VII, including in vivo of antisense control

The oligonucleotides listed in Table 97 below were tested in a non-terminal, dose escalation study for antisense inhibition of factor VII in monkeys.

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Each of the non-naive monkeys was injected subcutaneously with the oligonucleotides listed in Table 97 or PBS at escalating doses on days 0, 15, and 29. Each treatment group consisted of 4 males and 1 female. Before the first dose and at various time points thereafter, blood draws were performed to measure plasma factor VII protein levels. Factor VII protein levels were measured by ELISA. The results presented in Table 98 are the mean values of each treatment group relative to the mean values of the PBS group at baseline (BL), the measurement being taken immediately before the first dose. As demonstrated in Table 98, treatment with antisense oligonucleotides reduced Factor VII expression levels in a dose-dependent manner, and oligonucleotides with GalNAc conjugates were found in monkeys compared to oligonucleotides without GalNAc conjugates. significantly stronger.

Table 97

Oligonucleotides targeting factor VII

A legend to Table 97 can be found in Example 74. The structure of “GalNAc ₃ -10 _a ” is shown in Example 46.

Table 98

Factor VII Plasma Protein Levels

Example 92: GalNAc ₃ a conjugate containing Apo - CIII targeted antisense In primary hepatocytes by oligonucleotides antisense control

Primary mouse hepatocytes were seeded in 96-well plates at 15,000 cells/well and the oligonucleotides listed in Table 99 targeting mouse ApoC-III were 0.46, 1.37, 4.12, or 12.35, 37.04, 111.11, or 333.33 was added at nM or 1.00 μm. After incubation with oligonucleotides for 24 h, cells were lysed and total RNA was purified using RNeasy (Qiagen). ApoC-III mRNA levels were measured using real-time PCR and RiboGreen® RNA Quantification Reagent (Molecular Probes, Inc.) according to standard protocols. IC ₅₀ values were measured using Prism 4 software (GraphPad). These results indicate that, regardless of whether the cleavable moiety was a phosphodiester or a phosphodiester-linked deoxyadenosine, the oligonucleotides comprising the GalNAc conjugate were significantly stronger than the parent oligonucleotides without the conjugate. show

Table 99

Mice among mouse primary hepatocytes APOC Inhibition of -III expression

The structure of GalNAc ₃ -1 _a is already shown in Example 9, the structure of GalNAc ₃ -3 _a is shown in Example 39, _{the structure of GalNAc 3-7 a} is shown in Example 48, and GalNAc ₃ -10 the structure of _a may appear in example 46, the structure of _a GalNAc ₃ -13 may appear in example 62, and the structure of GalNAc ₃ -19 _a are set forth in example 70.

Example 93: Mixed Wings and 5'- GalNAc ₃ a conjugate containing SRB by oligonucleotides targeting -1 in vivo of antisense control

The oligonucleotides listed in Table 100 were tested in a dose-dependent study for antisense inhibition of SRB-1 in mice.

Table 100

SRB Modified to target -1 ASO

The structure of GalNAc ₃ -3 _a was already shown in Example 39, and _{the structure of GalNAc 3} -7a was already shown in Example 48. subscript: "e" represents a 2'-MOE modified nucleoside; "d" represents β-D-2'-deoxyribonucleoside;"k" represents 6'- (S) -CH ₃ bicyclic nucleoside (cEt); "s" represents a phosphorothioate internucleoside linkage (PS); "o" represents a phosphodiester internucleoside linkage (PO). The superscript "m" represents 5-methylcytosine.

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6-8 week old C57BL/6 mice (Jackson Laboratory, Bar Harbor, ME) were injected once subcutaneously with the oligonucleotides listed in Table 100 or saline at the doses shown below. Each treatment group consisted of 4 animals. Mice were sacrificed 72 hours after the last dose. Liver SRB-1 mRNA levels were measured using real-time PCR. SRB-1 mRNA levels were normalized to cyclophilin mRNA levels according to standard protocols. Results are presented as average percent of SRB-1 mRNA levels for each treatment group relative to the saline control group. As demonstrated in Table 101, treatment with antisense oligonucleotides reduced SRB-1 mRNA levels in a dose-dependent manner, and gapmer oligonucleotides with wings that contained GalNAc conjugates and were either complete cEt or mixed sugar modifications. was significantly stronger than the parent oligonucleotide without the conjugate and containing the complete cEt modified wing.

Body weight, liver transaminase, total bilirubin, and BUN were also measured and the mean values for each treatment group are shown in Table 101. Body weight is shown as mean percent body weight relative to baseline body weight (% BL) measured immediately prior to oligonucleotide dose.

Table 101

SRB -One mRNA , ALT, AST , BUN, and total bilirubin levels and body weight

Example 94: 2' -sugar modifications and 5'- GalNAc ₃ a conjugate containing SRB by oligonucleotides targeting -1 in vivo of antisense control

The oligonucleotides listed in Table 102 were tested in a dose-dependent study for antisense inhibition of SRB-1 in mice.

Table 102

SRB Modified to target -1 ASO

The subscript "m" denotes a 2'-0-methyl modified nucleoside. See Example 74 for complete table legend. The structure of GalNAc ₃ -3 _a was already shown in Example 39, and _{the structure of GalNAc 3} -7a was already shown in Example 48.

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This study was completed using the protocol described in Example 93. The results are shown in Table 103 below and show that both the 2'-MOE and 2'-OMe modified oligonucleotides comprising the GalNAc conjugate were significantly stronger than the respective parent oligonucleotides without the conjugate. Results of body weight, liver transaminase, total bilirubin, and BUN measurements indicated that all compounds were tolerated.

Table 103

SRB -One mRNA

Example 95: bicyclic nucleoside and 5'- GalNAc ₃ a conjugate containing SRB by oligonucleotides targeting -1 in vivo of antisense control

The oligonucleotides listed in Table 104 were tested in a dose-dependent study for antisense inhibition of SRB-1 in mice.

Table 104

SRB Modified to target -1 ASO

The subscript “g” denotes a fluoro-HNA nucleoside and the subscript “l” denotes a locked nucleoside comprising a _{2′-O—CH 2 -4′ bridge.} See Example 74 table legend for other abbreviations. The structure of GalNAc ₃ -1 _a was already shown in Example 9, the structure of GalNAc ₃ -3 _a was already shown in Example 39, and _{the structure of GalNAc 3} -7a was already shown in Example 48.

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This study was completed using the protocol described in Example 93. The results are shown in Table 105 below and show that the oligonucleotides containing the GalNAc conjugate and various bicyclic nucleoside modifications were significantly stronger than the parent oligonucleotide without the conjugate and containing the bicyclic nucleoside modifications. Moreover, the oligonucleotides containing the GalNAc conjugate and fluoro-HNA modification were significantly stronger than the parent oligonucleotide without the conjugate and containing the fluoro-HNA modification. Results of body weight, liver transaminase, total bilirubin, and BUN measurements indicated that all compounds were tolerated.

Table 105

SRB -One mRNA , ALT, AST , BUN, and total bilirubin levels and body weight

Example 96: GalNAc ₃ conjugate containing a group antisense Plasma protein binding of oligonucleotides

The oligonucleotides listed in Table 70 targeting ApoC-III and the oligonucleotides in Table 106 targeting apo(a) were tested in an ultrafiltration assay to assess plasma protein binding.

Table 106

apo (a) Targeted modified oligonucleotides

See Example 74 for table legend. The structure of “GalNAc ₃ -7a” is already shown in Example 48.

Ultrafree-MC ultrafiltration apparatus (30,000 NMWL, low-binding regenerated cellulose membrane, Millipore, Bedford, MA) was pre-conditioned with 300 μL of 0.5% Tween 80 and centrifuged at 2000 g for 10 min, then Pre-conditioned with a 300 μg/mL solution of 300 μL of control oligonucleotide in H _{2 O and centrifuged at 2000 g for 16 min.} To assess non-specific binding to the filter of each test oligonucleotide from Tables 70 and 106 to be used in this study, a 250 ng/mL solution of 300 μL of oligonucleotide in _{H 2 O was pre-conditioned at pH 7.4.} filtered and centrifuged at 2000 g for 16 min. Unfiltered and filtered samples were analyzed in an ELISA assay to determine oligonucleotide concentrations. Three replicates were used to obtain the average concentration for each sample. The average concentration of the filtered sample over the unfiltered sample is used to determine the percentage of oligonucleotide recovered through the filter in the absence of plasma (% recovery).

Frozen whole plasma samples collected in K3-EDTA from normal, drug-free human volunteers, cynomolgus monkeys, and CD-1 mice were purchased from Bioreclamation LLC (Westbury, NY). Test oligonucleotides were added to 1.2 mL aliquots of plasma at two concentrations (5 and 150 μg/mL). An aliquot (300 μL) of each spiked plasma sample was placed in a pre-conditioned filter device and incubated at 37° for 30 min, immediately followed by centrifugation at 2000 g for 16 min. Aliquots of filtered and unfiltered spiked plasma samples were analyzed by ELISA to determine oligonucleotide concentrations in each sample. Three replicates per concentration were used to determine the average percentage of bound and unbound oligonucleotides in each sample. The average concentration of the filtered sample relative to the concentration of the unfiltered sample is used to determine the percentage of oligonucleotides in plasma (% unbound) that are not bound to plasma proteins. The final unbound oligonucleotide value is corrected for non-specific binding by dividing the % unbound by the % recovery for each oligonucleotide. The final % bound oligonucleotide value is determined by subtracting the final % unbound value from 100. The results are shown in Table 107 for the concentration tests (5 and 150 μg/mL) of the two oligonucleotides tested in the plasma of each species. These results show that the GalNAc conjugate group has no significant effect on plasma protein binding. Moreover, oligonucleotides with both perfect PS internucleoside linkages and mixed PO/PS linkages bind plasma proteins, and those with perfect PS linkages to some extent greater than those with mixed PO/PS linkages. Binds to plasma proteins.

Table 107

to plasma proteins combined Percentage of modified oligonucleotides

Example 97: GalNAc ₃ conjugate containing a group TTR Targeted modified oligonucleotides

The oligonucleotides shown in Table 108 containing the GalNAc conjugate were designed to target the TTR.

Table 108

TTR Targeted modified oligonucleotides

A legend for Table 108 can be found in Example 74. The structure of “GalNAc ₃ -1” is shown in Example 9. The structure of “GalNAc ₃ -3 _a " is shown in Example 39. The structure of "GalNAc ₃ -7 _a " is shown in Example 48. The structure of “GalNAc ₃ -10 _a " is shown in Example 46. The structure of "GalNAc ₃ -13 _a " is shown in Example 62. The structure of “GalNAc ₃ -19 _a ” is shown in Example 70.

Example 98: hPMBC in black GalNAc a conjugate Evaluation of the pro-inflammatory effect of oligonucleotides comprising

The oligonucleotides listed in Table 109 were tested for pro-inflammatory effects in the hPMBC assay as described in Examples 23 and 24. (For description of oligonucleotides, see Tables 30, 83, 95, and 108). ISIS 353512 is a high responder used as a positive control, and other oligonucleotides are listed in Tables 83, 95, and 108. These results, shown in Table 109, were obtained using blood from one volunteer donor. These results show that oligonucleotides with mixed PO/PS internucleoside linkages produced significantly lower pro-inflammatory responses compared to the same oligonucleotides with perfect PS linkages. Moreover, the GalNAc conjugate group had no significant effect in this assay.

Table 109

Example 99: asialoglycoprotein for the receptor GalNAc a conjugate Binding affinity of oligonucleotides comprising

The binding affinity of the oligonucleotides listed in Table 110 to the asialoglycoprotein receptor (see Table 76 for a description of oligonucleotides) was tested in a competitive receptor binding assay. The competitor ligand, α1-acid glycoprotein (AGP), was incubated with 1 U neuraminidase-agarose in 50 mM sodium acetate buffer (pH 5) for 16 h at 37° C., >90% desialylation was Confirmed by sialic acid assay or size exclusion chromatography (SEC). Iodine monochloride is used the AGP has iodide according to the procedure due Atsma (Note:. J Lipid Res 1991 Jan; 32 (1): 173-81). In this method, desialylated α1-acid glycoprotein (de-AGP) was added to 10 mM iodine chloride, Na125I, and 1 M glycine in 0.25 M NaOH. After incubation for 10 min at room temperature, ¹²⁵ I -labeled de-AGP was isolated from ^{free 125} I by concentrating the mixture twice using a 3 KDMWCO spin column. Proteins were tested for labeling efficiency and purity on an Agilent SEC-3 column (7.8x300 mm) and an HPLC system equipped with a β-RAM counter. ^{Competition experiments using various GalNAc-clusters containing 125} I-labeled de-AGP and ASO were performed as follows. Human HepG2 cells (10 ⁶ cells/ml) were plated on 6-well plates in 2 mL of appropriate growth medium. MEM medium supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine and 10 mM HEPES was used. Cells were incubated with _{5% and 10% CO 2} respectively at 37° C. for 16-20 hours. Cells were washed with FBS-free medium before experiments. Cells were transferred to GalNAc-clusters containing 1 ml competition mixture containing appropriate growth medium with 2% FBS, 10 ^-8 M ¹²⁵ I-labeled de-AGP and ^{ASO at concentrations ranging from 10 -11} to 10 ^-5. Incubated at 37˚C for 30 min. Non-specific binding was determined in the presence ^{of 10 -2 M GalNAc sugar.} Cells were washed twice with FBS-free medium ^{to remove unbound 125} I-labeled de-AGP and competitor GalNAc ASO. Cells were lysed using Qiagen's RLT buffer containing 1% β-mercaptoethanol. Lysates were transferred to round bottom assay tubes after a short 10 min freeze/thaw cycle and assayed on a γ-counter. After subtracting non-specific binding, the ¹²⁵ I protein count was divided by the value of the lowest GalNAc-ASO concentration count. Inhibition curves were fitted according to the single site competitive binding equation using a nonlinear regression algorithm to calculate the _{binding affinity (K D's).}

The results in Table 110 were obtained from experiments performed on 5 different days. The results for the oligonucleotides indicated by the superscript “a” are the average of experimental runs on two different days. These results show that oligonucleotides containing a GalNAc conjugate group on the 5'-end have 1.5 to 16-fold greater affinity than oligonucleotides containing a GalNAc conjugate group on the 3'-terminus of human HepG2 cells. shows that it binds to the asialoglycoprotein receptor.

Table 110

asialoglycoprotein Receptor Binding Assay Results

Example 100: in vivo at apo(a) cast targeted GalNAc conjugate by an oligonucleotide comprising a group in vivo of antisense control

The oligonucleotides listed in Table 111a below were tested in a single dose study for duration of action in mice.

Table 111a

apo (a) modified to target ASO

The structure of “GalNAc ₃ -7 _a ” is shown in Example 48.

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Each of the female transgenic mice expressing human spores (a) was injected subcutaneously once per week, for a total of 6 doses, with the oligonucleotides and doses listed in Table 111b or PBS. Each treatment group consisted of 3 animals. Blood was drawn the day before dosing and baseline levels of apo(a) protein were determined in plasma and at 72 hours, 1 week, and 2 weeks following the first dose. Additional blood draws will occur at 3 weeks, 4 weeks, 5 weeks, and 6 weeks following the first dose. Plasma apo(a) protein levels were measured using ELISA. The results in Table 111b are presented as the mean percent of plasma apo(a) protein levels for each treatment group, normalized to baseline levels (% BL), indicating that oligonucleotides comprising the GalNAc conjugate group (a) shows a strong reduction in expression. This strong effect was observed for oligonucleotides containing complete PS internucleoside linkages and oligonucleotides containing mixed PO and PS linkages.

Table 111b

spore (a) plasma protein levels

Example 101: stable a moiety connected through GalNAc by oligonucleotides comprising clusters antisense control

The oligonucleotides listed in Table 112 were tested in vivo for inhibition of mouse APOC-III expression. Each of the C57Bl/6 mice was injected subcutaneously once with the oligonucleotides listed in Table 112 or PBS. Each treatment group consisted of 4 animals. Each mouse treated with ISIS 440670 received a dose of 2, 6, 20, or 60 mg/kg. Each mouse treated with ISIS 680772 or 696847 received 0.6, 2, 6, or 20 mg/kg. The GalNAc conjugate group of ISIS 696847 is linked via a stable moiety, phosphorothioate linkage instead of a readily cleavable phosphodiester containing linkage. Animals were sacrificed 72 hours after dosing. Liver APOC-III mRNA levels were measured using real-time PCR. APOC-III mRNA levels were normalized to cyclophilin mRNA levels according to standard protocols. Results are provided in Table 112 as the average percent of APOC-III mRNA levels for each treatment group relative to the saline control group. These results show that the oligonucleotides containing the GalNAc conjugate group were significantly stronger than the oligonucleotides without the conjugate group. Moreover, oligonucleotides comprising a GalNAc conjugate group linked to the oligonucleotide via a cleavable moiety (ISIS 680772) are more oligonucleotides than those comprising a GalNAc conjugate group linked to the oligonucleotide via a stable moiety (ISIS 696847). stronger

Table 112

mouse APOC Modified oligonucleotides targeting -III

The structure of “GalNAc ₃ -7 _a ” is shown in Example 48.

Example 102: GalNAc a conjugate containing antisense Distribution of oligonucleotides in the liver

The liver distribution of ISIS 353382 without GalNAc conjugate (Ref. Table 36) and ISIS 655861 with GalNAc conjugate (Ref. Table 36) was evaluated. Male Balb/c mice were injected subcutaneously once with either ISIS 353382 or 655861 at the doses listed in Table 113. Each treatment group consisted of 3 animals with the exception of the 18 mg/kg group for ISIS 655861 which consisted of 2 animals. Animals were sacrificed 48 hours after dosing to determine the liver distribution of oligonucleotides. To determine the number of antisense oligonucleotide molecules per cell, a ruthenium (II) tris-bipyridine tag (MSD TAG, Medium Discovery) is conjugated to an oligonucleotide probe used to detect antisense oligonucleotides. Results presented in Table 113 are average concentrations of oligonucleotides for each treatment group in millions of oligonucleotide molecules per cell. These results show that, at equivalent doses, oligonucleotides with GalNAc conjugates are present in higher concentrations in whole liver and hepatocytes than oligonucleotides without GalNAc conjugates. Moreover, the oligonucleotides comprising the GalNAc conjugate were present at lower concentrations in non-parenchymal liver cells than the oligonucleotides without the GalNAc conjugate. And the concentration of ISIS 655861 in hepatocytes and non-parenchymal liver cells was similar for cells, but the liver is approximately 80% hepatocytes by volume. Thus, the majority of ISIS 655861 oligonucleotides present in the liver were found in hepatocytes, whereas the majority ISIS 353382 oligonucleotides present in the liver were found in non-parenchymal liver cells.

Table 113

Example 103: GalNAc ₃ a conjugate containing APOC of oligonucleotides targeting -III in vivo duration of action

The oligonucleotides listed in Table 114 below were tested in a single dose study for duration of action in mice.

Table 114

APOC Modified to target -III ASO

The structure of GalNAc ₃ -3 _a is already shown in Example 39, and GalNAc ₃ -19 _a is shown in Example 70.

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Each of the female transgenic mice expressing human APOC-III was injected subcutaneously once with the oligonucleotides listed in Table 114 or PBS. Each treatment group consisted of 3 animals. Blood was drawn prior to dosing and measured at baseline and on days 3, 7, 14, 21, 28, 35, and 42 following dosing. Plasma triglyceride and APOC-III protein levels were measured as described in Example 20. Results in Table 115 are presented as mean percentages of plasma triglyceride and APOC-III levels for each treatment group, normalized to baseline levels. A comparison between the results of Table 71 of Example 79 and the results of Table 115 below shows that an oligonucleotide comprising a mixture of phosphodiester and phosphorothioate internucleoside linkages is a phosphorothioate internucleoside linkage. showed an increased duration of action than equivalent oligonucleotides containing only

Table 115

Plasma in transgenic mice triglycerides and APOC -III protein level

Example 104: 5' - GalNAc ₂ a conjugate Synthesis of oligonucleotides comprising

Compound 120 is commercially available and the synthesis of compound 126 is described in Example 49. Compound 120 (1 g, 2.89 mmol) and HBTU (0.39 g, 2.89 mmol) and HOBt (1.64 g, 4.33 mmol) were dissolved in DMF (10 mL. N , N- N,N-diisopropylethylamine 1.75 mL, 10.1 mmol) was added. After about 5 minutes, aminohexanoic acid benzyl ester (1.36 g, 3.46 mmol) was added to the reaction. After 3 h, the reaction mixture was poured into 100 mL of 1 M NaHSO4 and extracted with 2 x 50 mL ethyl acetate. The organic layers were combined and _{washed with 3 x 40 mL saturated NaHCO 3} and 2 x brine _{, dried over Na 2} SO ₄ , filtered and concentrated. The product was purified by silica gel column chromatography (DCM:EA:Hex, 1:1:1) to obtain compound 231. LCMS and NMR were consistent with the structure. Compound 231 (1.34 g, 2.438 mmol) was dissolved in dichloromethane (10 mL) and trifluoroacetic acid (10 mL) was added. After stirring at room temperature for 2 h, the reaction mixture was concentrated under reduced pressure and co-evaporated with toluene (3 x 10 mL). The residue was dried under reduced pressure to give compound 232 as a trifluoroacetate salt. The synthesis of compound 166 is described in Example 54. Compound 166 (3.39 g, 5.40 mmol) was dissolved in DMF (3 mL). A solution of compound 232 (1.3 g, 2.25 mmol) was dissolved in DMF (3 mL) and N , N -diisopropylethylamine (1.55 mL) was added. The reaction was stirred at room temperature for 30 min, then poured into water (80 mL) and the aqueous layer was extracted with EtOAc (2x100 mL). The organic phase was separated _{and washed with saturated aqueous NaHCO 3} (3 x 80 mL), 1 M NaHSO ₄ (3 x 80 mL) and brine (2 x 80 mL), then dried (Na ₂ SO ₄ ) and filtered. and concentrated. The residue was purified by silica gel column chromatography to obtain compound 233. LCMS and NMR were consistent with the structure. Compound 233 (0.59 g, 0.48 mmol) was dissolved in methanol (2.2 mL) and ethyl acetate (2.2 mL). Palladium on carbon (10 wt % Pd/C, wet, 0.07 g) was added and the reaction mixture was stirred under a hydrogen atmosphere for 3 h. The reaction mixture was filtered through a pad of celite and concentrated to give the carboxylic acid. Carboxylic acid (1.32 g, 1.15 mmol, cluster free acid) was dissolved in DMF (3.2 mL). To this were added N , N -diisopropylethylamine (0.3 mL, 1.73 mmol) and PFPTFA (0.30 mL, 1.73 mmol). After stirring at room temperature for 30 min, the reaction mixture was poured into water (40 mL) and extracted with EtOAc (2 x 50 mL). Standard workup was completed as described above to yield compound 234. LCMS and NMR were consistent with the structure. Oligonucleotide 235 was prepared using the general procedure described in Example 46. The GalNAc ₂ cluster portion of the conjugate group GalNAc _{2 -24 (GalNAc 2} -24 _a ) can be combined with any cleavable moiety present on the oligonucleotide to provide a variety of conjugate groups. The structure of GalNAc ₂ -24 (GalNAc ₂ -24 _a -CM) is as follows:

Example 105: GalNAc _One -25 a conjugate Synthesis of oligonucleotides comprising

The synthesis of compound 166 is described in Example 54. Oligonucleotide 236 was prepared using the general procedure described in Example 46.

Alternatively, oligonucleotide 236 was synthesized using the scheme shown below, and compound 238 was used to form oligonucleotide 236 using the procedure described in Example 10.

GalNAc ₁ cluster portion of the conjugate group _{GalNAc 1 -25 (GalNAc 1 -25 a} ) is raised in combination with any cuttable moiety present on the oligonucleotide and can provide a variety of conjugate groups. The structure of GalNAc ₁ -25 (GalNAc ₁ -25 _a -CM) is as follows:

Example 106: 5' - GalNAc ₂ or 5'- GalNAc ₃ a conjugate containing SRB by oligonucleotides targeting -1 in vivo antisense control

The oligonucleotides listed in Tables 116 and 117 were tested in a dose-dependent study for antisense inhibition of SRB-1 in mice.

cure

ISIS number 440762 at 2, 7, or 20 mg/kg to 6 week old, male C57BL/6 mice (Jackson Laboratory, Bar Harbor, ME); or 0.2, 0.6, 2, 6, or 20 mg/kg of ISIS No. 686221, 686222, or 708561; or saline was administered once subcutaneously. Each treatment group consisted of 4 animals. Mice were sacrificed 72 hours after the last dose. Liver SRB-1 mRNA levels were measured using real-time PCR. SRB-1 mRNA levels were normalized to cyclophilin mRNA levels according to standard protocols. Antisense oligonucleotides reduced SRB-1 mRNA levels in a dose-dependent manner, and ED ₅₀ results are provided in Tables 116 and 117. Previous studies have shown that trivalent GalNAc-conjugated oligonucleotides were significantly stronger than bivalent GalNAc-conjugated oligonucleotides, which in turn were significantly stronger from monovalent GalNAc-conjugated oligonucleotides ( see , for example, Khorev et al., Bioorg . & Med . Chem ., Vol. 16, 5216-5231 (2008)), treatment with antisense oligonucleotides comprising monovalent, divalent, and trivalent GalNAc clusters. reduced SRB-1 mRNA levels with similar efficacy as shown in Tables 116 and 117.

Table 116

SRB Modified oligonucleotides targeting -1

See Example 93 for table legend. The structure of GalNAc ₃ -13a has already been shown in Example 62, the structure of GalNAc -24a ₂ is shown in Example 104.

Table 117

SRB Modified oligonucleotides targeting -1

See Example 93 for table legend. GalNAc has the structure of ₁ -25a shown in Example 105.

The concentrations of oligonucleotides in Tables 116 and 117 in the liver were also assessed using the procedure described in Example 75. Results shown in Tables 117a and 117b below are the mean total antisense oligonucleotide tissue levels for each treatment group, as measured by UV in μg oligonucleotides/gram of liver tissue. These results show that oligonucleotides comprising the GalNAc conjugate group accumulate in the liver at significantly higher levels than the same dose of oligonucleotides without the GalNAc conjugate group. Moreover, antisense oligonucleotides comprising one, two or three GalNAc ligands in all of their respective conjugate groups accumulate in the liver to similar levels. These results are surprising in view of the Khorev et al references cited above and are consistent with the activity data shown in Tables 116 and 117 above.

Table 117a

GalNAc ₂ or GalNAc ₃ conjugate Liver concentration of oligonucleotides comprising groups

Table 117b

GalNAc _One conjugate Liver concentration of oligonucleotides comprising groups

Example 107: GalNAc _One -26 or GalNAc _One -27 a conjugate Synthesis of oligonucleotides comprising

Oligonucleotide 239 is synthesized via coupling of compound 47 (Reference Example 15) to acid 64 (Reference Example 32) using HBTU and DIEA in DMF. The resulting amide containing compound is phosphitylated and then added to the 5'-end of the oligonucleotide using the procedure described in Example 10. GalNAc ₁ cluster portion of the conjugate group _{GalNAc 1 -26 (GalNAc 1 -26 a} ) is raised in combination with any cuttable moiety present on the oligonucleotide and can provide a variety of conjugate groups. The structure of GalNAc ₁ -26 (GalNAc ₁ -26 _a -CM) is as follows:

.

.

To add a GalNAc ₁ conjugate group to the 3'-end of the oligonucleotide, the amide formed from the reaction of compounds 47 and 64 is added to the solid support using the procedure described in Example 7. Oligonucleotide synthesis is then completed using the procedure described in Example 9 to form oligonucleotide 240.

Konju GalNAc cluster ₁ of the gate group _{GalNAc 1 -27 (GalNAc 1 -27 a} ) is raised in combination with any cuttable moiety present on the oligonucleotide and can provide a variety of konju gate group. The structure of GalNAc ₁ -27 (GalNAc ₁ -27 _a -CM) is as follows:

Example 108: in vivo at apo(a) cast targeted GalNAc conjugate by an oligonucleotide comprising a group in vivo of antisense control

The oligonucleotides listed in Table 118 below were tested in a single dose study in mice.

Table 118

apo (a) modified to target ASO

The structure of “GalNAc ₃ -7 _a ” is shown in Example 48.

cure

Each of the male transgenic mice expressing human spores (a) was injected subcutaneously once with oligonucleotides and at the doses listed in Table 119 or PBS. Each treatment group consisted of 4 animals. Blood was drawn the day before dosing and one week following the first dose to determine baseline levels of apo(a) protein in plasma. Additional blood draws will occur weekly for approximately 8 weeks. Plasma apo(a) protein levels were measured using ELISA. Results in Table 119 are presented as the mean percent of plasma apo(a) protein levels for each treatment group, normalized to baseline levels (% BL). These results show that antisense oligonucleotides reduce apo(a) protein expression. Moreover, the oligonucleotides containing the GalNAc conjugate group showed a stronger reduction in apo(a) expression than the oligonucleotides without the conjugate group.

Table 119

Apo(a) plasma protein levels

Example 109: GalNAc _One -28 or GalNAc _One -29 Synthesis of oligonucleotides comprising conjugates

Oligonucleotide 241 is synthesized using a procedure similar to that described in Example 71 to form the phosphoramidite intermediate, and then the oligonucleotide is synthesized using the procedure described in Example 10. Conjugate group GalNAc ₁ cluster portion of _{GalNAc 1 -28 (GalNAc 1 -28 a} ) is raised in combination with any cuttable moiety present on the oligonucleotide and can provide a variety of conjugate groups. The structure of GalNAc ₁ -28 (GalNAc ₁ -28 _a -CM) is as follows:

To add a GalNAc ₁ conjugate group to the 3'-end of the oligonucleotide, a procedure similar to that described in Example 71 is used to form a hydroxyl intermediate, which is then used as a solid support using the procedure described in Example 7 is added to Oligonucleotide synthesis is then completed using the procedure described in Example 9 to form oligonucleotide 242.

GalNAc ₁ cluster portion of the conjugate group _{GalNAc 1 -29 (GalNAc 1 -29 a} ) is raised in combination with any cuttable moiety present on the oligonucleotide and can provide a variety of conjugate groups. The structure of GalNAc ₁ -29 (GalNAc ₁ -29 _a -CM) is as follows:

.

.

Example 110: GalNAc _One -30 Synthesis of oligonucleotides comprising conjugates

GalNAc ₁ -30 konju oligonucleotide comprising the nucleotide gate group 246 (where Y is O, S, substituted or unsubstituted C ₁ -C ₁₀ alkyl, amino, selected from substituted amino, azido, alkenyl, or alkynyl ) is synthesized as shown above. GalNAc ₁ cluster portion of the conjugate group _{GalNAc 1 -30 (GalNAc 1 -30 a} ) is raised in combination with any cuttable moiety present on the oligonucleotide and can provide a variety of conjugate groups. In certain embodiments, Y is part of a cleavable moiety. In certain embodiments, Y is part of a stable moiety and the cleavable moiety is present on the oligonucleotide. It is shown for the structure of the _"a GalNAc ₁ -30".

Example 111: GalNAc ₂ -31 or GalNAc ₂ Synthesis of oligonucleotides comprising -32 conjugates

Oligonucleotide 250 comprising a GalNAc ₂ -31 conjugate group, wherein Y is selected from O, S, substituted or unsubstituted C ₁ -C ₁₀ alkyl, amino, substituted amino, azido, alkenyl or alkynyl. ) is synthesized as shown above. The GalNAc ₂ cluster portion of the conjugate group GalNAc _{2 -31 (GalNAc 2} -31 _a ) can be combined with any cleavable moiety present on the oligonucleotide to provide a variety of conjugate groups. In certain embodiments, the Y-containing group directly adjacent to the 5'-end of the oligonucleotide is part of a cleavable moiety. In certain embodiments, the Y-containing group directly adjacent to the 5'-end of the oligonucleotide is part of the stable moiety and the cleavable moiety is present on the oligonucleotide. The structure of "GalNAc ₂ -31 _a " is shown below.

The synthesis of an oligonucleotide comprising a GalNAc _{2 -32 conjugate is shown below.}

Oligonucleotide 252 comprising a GalNAc ₂ -32 conjugate group, wherein Y is selected from O, S, substituted or unsubstituted C ₁ -C ₁₀ alkyl, amino, substituted amino, azido, alkenyl or alkynyl. ) is synthesized as shown above. The GalNAc ₂ cluster portion of the conjugate group GalNAc _{2 -32 (GalNAc 2} -32 _a ) can be combined with any cleavable moiety present on the oligonucleotide to provide a variety of conjugate groups. In certain embodiments, the Y-containing group directly adjacent to the 5'-end of the oligonucleotide is part of a cleavable moiety. In certain embodiments, the Y-containing group directly adjacent to the 5'-end of the oligonucleotide is part of the stable moiety and the cleavable moiety is present on the oligonucleotide. The structure of "GalNAc ₂ -32 _a " is shown below.

Example 112: GalNAc _One Synthesis of oligonucleotides comprising conjugates

Oligonucleotides in Table 120 targeting SRB-1 were synthesized with a group of _{GalNAc 1} conjugates to further test the efficacy of oligonucleotides comprising a group of conjugates containing one GalNAc ligand.

Table 120

Example 113: Antisense Oligonucleotides Targeting Angiopoietin-Like 3 and Containing a GalNAc Conjugate Group

The oligonucleotides in Table 121 were designed to target angiopoietin-like 3 (ANGPTL3).

Table 121

Example 114: By oligonucleotides comprising a group of GalNAc conjugates targeting human ANGPTL3 in vivo antisense inhibition of

Oligos listed in Table 122 (and listed in Table 121), expressing human ANGPTL3, intraperitoneally into 6 week old, male transgenic C57Bl/6 mice once per week at the doses shown below for a total of 2 doses. Nucleotides or PBS were injected. Each treatment group consisted of 4 animals. Mice were sacrificed 2 days after the last dose. ANGPTL3 liver mRNA levels were measured using real-time PCR and RiboGreen® RNA Quantification Reagent (Molecular Probes, Inc. Eugene, OR) according to standard protocols. The results below are presented as the average percent of ANGPTL3 mRNA levels in the liver for each treatment group normalized to the PBS control.

As demonstrated in Table 122, treatment with antisense oligonucleotides reduced ANGPTL3 liver mRNA levels in a dose-dependent manner, and oligonucleotides with GalNAc conjugates were significantly more potent than parental oligonucleotides without GalNAc conjugates. It was strong.

Table 122

ANGPTL3 liver mRNA levels

Liver alanine transaminase (ALT) levels were also measured at sacrifice using standard protocols. The results showed that none of the treatment groups had elevated ALT levels, indicating that the oligonucleotides were well tolerated.

Example 115: Using an oligonucleotide comprising a group of GalNAc conjugates targeting mouse ANGPTL3 in vivo antisense inhibition of

The oligonucleotides listed in Table 123 below were tested in a dose dependent study in mice.

Table 123

mouse ANGPTL3 modified to target ASO

The structure of “GalNAc ₃ -7 _a ” is shown in Example 48.

Low-density lipoprotein receptor knockout (LDLR-/-) mice were fed on a Western diet for one week prior to intraperitoneal injection once a week with the oligonucleotides listed in Table 123 or PBS at the doses shown below. Each treatment group consisted of 5 animals. Blood was drawn after the first dose was administered and 2 weeks after the first dose to determine baseline levels of triglycerides in plasma. The results in Table 124 are presented as the mean percent of plasma triglyceride levels for each treatment group, normalized to baseline levels (% BL), indicating that antisense oligonucleotides reduce triglyceride expression in a dose dependent manner. show that Moreover, the oligonucleotides containing the GalNAc conjugate group showed a stronger reduction in triglycerides than the oligonucleotides without the conjugate group.

Table 124

Plasma Triglyceride (TG) Levels

Example 116: Antisense inhibition of human angiopoietin-like 3 by MOE gapmers in Hep3B cells

Antisense oligonucleotides were designed to target angiopoietin-like 3 (ANGPTL3) nucleic acid and tested for their effect on ANGPTL3 mRNA in vitro. The antisense oligonucleotides were tested for potency in a series of parallel experiments with similar culture conditions. The results for each experiment are presented in a separate table shown below. Hep3B cells cultured at a density of 20,000 cells per well were transfected with 4,500 nM antisense oligonucleotides using electroporation. After a treatment period of approximately 24 hours, RNA was isolated from the cells and ANGPTL3 mRNA levels were measured by quantitative real-time PCR. Using the human primer probe set RTS3492_MGB (forward sequence CCGTGGAAGACCAATATAAAACAATT, designated herein as SEQ ID NO: 4; AGTCCTTCTGAGCTGATTTTCTATTTCT; reverse sequence, designated herein as SEQ ID NO: 5; probe sequence AACCAACAGCATAGTCAAATA, designated herein as SEQ ID NO: 6) mRNA levels were measured. ANGPTL3 mRNA levels were adjusted according to total RNA content as measured by Ribogreen®. Results are expressed as percent inhibition of ANGPTL3 relative to untreated control cells.

In the table below, the newly designed chimeric antisense oligonucleotides were designed as 5-10-5 MOE gapmers. 5-10-5 MOE gapmers are 20 nucleotides in length, wherein the central gap segment comprises 10 2' deoxynucleosides and wing segments in the 5' direction and 3' direction comprising 5 nucleotides each is flanked by Each nucleoside in the 5' wing segment and each nucleoside in the 3' wing segment has a 2'-MOE modification. The internucleoside linkages throughout each gapmer are phosphorothioate (P=S) linkages. All cytosine residues throughout each gapmer are 5-methylcytosine. "Initiation site" refers to the 5'-most nucleoside to which the gapmer is targeted within the human gene sequence. "Stop site" refers to the 3'-most nucleoside to which the gapmer is targeted within the human gene sequence. Each of the gapmers listed in the table below is a human ANGPTL3 mRNA (GenBank Accession No. NM_014495.2) or human ANGPTL3 genomic sequence (assigned herein as SEQ ID NO: 2; 33032001 to 33046000 as disclosed herein) gene bank accession number NT_032977.9) excised from 'n/a' means that the antisense oligonucleotide does not target a specific gene sequence with 100% complementarity.

Table 125

Inhibition of ANGPTL3 mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 1 and 2

Table 126

Inhibition of ANGPTL3 mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 1 and 2

Table 127

Inhibition of ANGPTL3 mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 1 and 2

Table 128

Inhibition of ANGPTL3 mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 1 and 2

Table 129

Inhibition of ANGPTL3 mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 1 and 2

Table 130

Inhibition of ANGPTL3 mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 1 and 2

Table 131

Inhibition of ANGPTL3 mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 1 and 2

Table 132

Inhibition of ANGPTL3 mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 1 and 2

Table 133

Inhibition of ANGPTL3 mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 1 and 2

Table 134

Inhibition of ANGPTL3 mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 1 and 2

Table 135

Inhibition of ANGPTL3 mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 1 and 2

Table 136

Inhibition of ANGPTL3 mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 1 and 2

Table 137

Inhibition of ANGPTL3 mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 1 and 2

Table 138

Inhibition of ANGPTL3 mRNA by 5-10-5 MOE gapmers targeting SEQ ID NOs: 1 and 2

Example 117: MOE to gapmer by Hep3B within the cell human of ANGPTL3 Dose-dependent antisense control

5-10-5 MOE gapmers from the study described above that showed significant in vitro inhibition of ANGPTL3 mRNA were selected and tested at various doses in Hep3B cells. Cells were plated at a density of 20,000 cells per well and transfected using electroporation with antisense oligonucleotides at concentrations of 0.75 μM, 1.50 μM, 3.00 μM, 6.00 μM and 12.00 μM, as indicated in the table below. After a treatment period of approximately 16 hours, RNA was isolated from the cells and ANGPTL3 mRNA levels were measured by quantitative real-time PCR. mRNA levels were measured using the human primer probe set RTS3492_MGB. ANGPTL3 mRNA levels were adjusted according to total RNA content as measured by RiboGreen®. Results are expressed as percent inhibition of ANGPTL3 relative to untreated control cells.

The half maximal inhibitory concentration (IC ₅₀ ) of each oligonucleotide is also presented in the table below. ANGPTL3 mRNA levels were significantly reduced in a dose-dependent manner in antisense oligonucleotide-treated cells.

Table 139

Example 118: Dose-dependent antisense inhibition of human ANGPTL3 in Hep3B cells by MOE gapmers

5-10-5 MOE gapmers from the study described above that showed significant in vitro inhibition of ANGPTL3 mRNA were selected and tested at various doses in Hep3B cells. Cells were plated at a density of 20,000 cells per well and transfected with antisense oligonucleotides at concentrations of 0.813 μM, 1.625 μM, 3.25 μM, 6.50 μM and 13.00 μM, as indicated in the table below. After a treatment period of approximately 16 hours, RNA was isolated from the cells and ANGPTL3 mRNA levels were measured by quantitative real-time PCR. mRNA levels were measured using the human primer probe set RTS3492_MGB. ANGPTL3 mRNA levels were adjusted according to total RNA content as measured by ^{RiboGreen ®.} Results are expressed as percent inhibition of ANGPTL3 relative to untreated control cells.

The half maximal inhibitory concentration (IC ₅₀ ) of each oligonucleotide is also presented in the table below. ANGPTL3 mRNA levels were significantly reduced in a dose-dependent manner in antisense oligonucleotide-treated cells.

Table 140

Table 141

Table 142

Table 143

Table 144

Table 145

Table 146

Table 147

Example 119: Antisense inhibition of human ANGPTL3 in Hep3B cells by deoxy, MOE and (S)-cEt gapmers

Additional antisense oligonucleotides were designed to target the ANGPTL3 nucleic acid and tested for their effect on ANGPTL3 mRNA in vitro. Hep3B cells cultured at a density of 20,000 cells per well were transfected using electroporation with 4,500 nM antisense oligonucleotides. After a treatment period of approximately 24 hours, RNA was isolated from the cells and ANGPTL3 mRNA levels were measured by quantitative real-time PCR. mRNA levels were measured using the human primer probe set RTS3492_MGB. ANGPTL3 mRNA levels were adjusted according to the total RNA content as measured by ^{RiboGreen ®.} Results are expressed as percent inhibition of ANGPTL3 relative to untreated control cells.

Newly designed chimeric antisense oligonucleotides in the table below were designed as deoxy, MOE, and (S)-cEt oligonucleotides. Deoxy, MOE and (S)-cEt oligonucleotides are 16 nucleotides in length, wherein the nucleosides have MOE sugar modifications, (S)-cEt sugar modifications, or deoxy sugar residues. The sugar modification of each antisense oligonucleotide is described as 'eek-d10-kke', where 'k' represents the (S)-cEt sugar modification; 'd' represents deoxyribose; The number indicates the number of residues per deoxyribose; 'e' represents the modification per MOE. The internucleoside linkage throughout each oligonucleotide is a phosphorothioate (P=S) linkage. All cytosine residues throughout each oligonucleotide are 5-methylcytosine. "Initiation site" refers to the 5'-most nucleoside to which the oligonucleotide is targeted within a human gene sequence. "Stop site" refers to the 3'-most nucleoside to which the oligonucleotide is targeted within a human gene sequence. Each of the oligonucleotides listed in the table below is a human ANGPTL3 mRNA (GenBank Accession No. NM_014495.2) or human ANGPTL3 genomic sequence (assigned herein as SEQ ID NO: 2; 33032001 to SEQ ID NO: 1 as disclosed herein) gene bank accession number NT_032977.9) cleaved from 33046000). 'n/a' means that the antisense oligonucleotide does not target a specific gene sequence with 100% complementarity.

Table 148

Inhibition of ANGPTL3 mRNA by deoxy, MOE and cEt oligonucleotides targeting SEQ ID NOs: 1 and 2

Table 149

Inhibition of ANGPTL3 mRNA by deoxy, MOE and (S)-cEt gapmers targeting SEQ ID NOs: 1 and 2

Example 120: Antisense inhibition of human ANGPTL3 in Hep3B cells by deoxy, MOE and (S)-cEt gapmers

Additional antisense oligonucleotides were designed to target the ANGPTL3 nucleic acid and tested for their effect on ANGPTL3 mRNA in vitro. The 5-10-5 MOE gapmers ISIS 337487 and ISIS 233717 were also included in the assay as benchmark oligonucleotides. Hep3B cells cultured at a density of 20,000 cells per well were transfected with 4,500 nM antisense oligonucleotides using electroporation. After a treatment period of approximately 24 hours, RNA was isolated from the cells and ANGPTL3 mRNA levels were measured by quantitative real-time PCR. mRNA levels were measured using the human primer probe set RTS3492_MGB. ANGPTL3 mRNA levels were adjusted according to total RNA content as measured by Ribogreen®. Results are expressed as percent inhibition of ANGPTL3 relative to untreated control cells.

Newly designed chimeric antisense oligonucleotides in the table below were designed with deoxy, MOE, and (S)-cEt oligonucleotides or 5-10-5 MOE gapmers. Deoxy, MOE and (S)-cEt oligonucleotides are 16 nucleotides in length, wherein the nucleoside has a MOE sugar modification, (S)-cEt sugar modification, or deoxy sugar residue. The sugar modification of each antisense oligonucleotide is described as 'eek-d10-kke', where 'k' is the (S)-cEt sugar modification; 'd' represents deoxyribose; The number indicates the number of residues per deoxyribose; 'e' represents the MOE modification. 5-10-5 MOE gapmers are 20 nucleotides in length, wherein the central gap segment comprises 10 2' deoxynucleosides and wing segments in the 5' direction and 3' direction comprising 5 nucleotides each is flanked by The internucleoside linkage throughout each oligonucleotide is a phosphorothioate (P=S) linkage. All cytosine residues throughout each oligonucleotide are 5-methylcytosine. "Initiation site" refers to the 5'-most nucleoside to which the oligonucleotide is targeted within a human gene sequence. "Stop site" refers to the 3'-most nucleoside to which the oligonucleotide is targeted within a human gene sequence. Each of the oligonucleotides listed in the table below is a human ANGPTL3 mRNA (GenBank Accession No. NM_014495.2) or human ANGPTL3 genomic sequence (assigned herein as SEQ ID NO: 2; 33032001 to SEQ ID NO: 1 as disclosed herein) gene bank accession number NT_032977.9) cleaved from 33046000). 'n/a' means that the antisense oligonucleotide does not target a specific gene sequence with 100% complementarity.

Table 150

Inhibition of ANGPTL3 mRNA by oligonucleotides targeting SEQ ID NOs: 1 and 2

Table 151

Inhibition of ANGPTL3 mRNA by oligonucleotides targeting SEQ ID NOs: 1 and 2

Table 152

Inhibition of ANGPTL3 mRNA by oligonucleotides targeting SEQ ID NOs: 1 and 2

Table 153

Inhibition of ANGPTL3 mRNA by oligonucleotides targeting SEQ ID NOs: 1 and 2

Table 154

Inhibition of ANGPTL3 mRNA by oligonucleotides targeting SEQ ID NOs: 1 and 2

Example 121: Antisense inhibition of human ANGPTL3 in Hep3B cells by MOE gapmers

Additional antisense oligonucleotides were designed to target the ANGPTL3 nucleic acid and tested for their effect on ANGPTL3 mRNA in vitro. Hep3B cells cultured at a density of 20,000 cells per well were transfected with 4,500 nM antisense oligonucleotides using electroporation. After a treatment period of approximately 24 hours, RNA was isolated from the cells and ANGPTL3 mRNA levels were measured by quantitative real-time PCR. mRNA levels were measured using the human primer probe set RTS3492_MGB. ANGPTL3 mRNA levels were adjusted according to total RNA content as measured by Ribogreen®. Results are expressed as percent inhibition of ANGPTL3 relative to untreated control cells.

In the table below, the newly designed chimeric antisense oligonucleotides were designated as 5-10-5 MOE or 3-10-4 MOE gapmers. 5-10-5 MOE gapmers are 20 nucleotides in length, wherein the central gap segment comprises 10 2' deoxynucleosides and wing segments in the 5' direction and 3' direction comprising 5 nucleotides each is flanked by The 3-10-4 MOE gapmer is 17 nucleotides in length, wherein the central gap segment contains 10 2' deoxynucleosides and contains 3 and 4 nucleotides in the 5' direction and 3' direction, respectively. flanked by wing segments at Each nucleoside in the 5' wing segment and each nucleoside in the 3' wing segment has a 2'-MOE modification. Each nucleoside in the 5' wing segment and each nucleoside in the 3' wing segment has a 2'-MOE modification. The internucleoside linkages throughout each gapmer are phosphorothioate (P=S) linkages. All cytosine residues throughout each gapmer are 5-methylcytosine. "Initiation site" refers to the 5'-most nucleoside to which the gapmer is targeted within the human gene sequence. "Stop site" refers to the 3'-most nucleoside to which the gapmer is targeted within the human gene sequence. Each of the gapmers listed in the table below is a human ANGPTL3 mRNA (GenBank Accession No. NM_014495.2) or human ANGPTL3 genomic sequence (assigned herein as SEQ ID NO: 2; 33032001 to 33046000 as disclosed herein) gene bank accession number NT_032977.9) excised from 'n/a' means that the antisense oligonucleotide does not target a specific gene sequence with 100% complementarity.

Table 155

Inhibition of ANGPTL3 mRNA by MOE gapmers targeting SEQ ID NOs: 1 and 2

Example 122: Dose-dependent antisense inhibition of human ANGPTL3 in Hep3B cells

Deoxy, MOE, and cEt oligonucleotides were selected from the studies described above showing significant in vitro inhibition of ANGPTL3 mRNA and tested at various doses in Hep3B cells. Both ISIS 233717 and ISIS 337847, 5-10-5 MOE gapmers were also included in this study. The antisense oligonucleotides were tested for potency in a series of parallel experiments with similar culture conditions. The results of each experiment are shown in a separate table below.

Cells were plated at a density of 20,000 cells per well and transfected using electroporation using antisense oligonucleotides at concentrations of 0.813 μM, 1.625 μM, 3.25 μM, 6.500 μM and 13.00 μM, as indicated in the table below. After a treatment period of approximately 16 hours, RNA was isolated from the cells and ANGPTL3 mRNA levels were measured by quantitative real-time PCR. mRNA levels were measured using the human primer probe set RTS3492_MGB. ANGPTL3 mRNA levels were adjusted according to total RNA content as measured by RiboGreen®. Results are expressed as percent inhibition of ANGPTL3 relative to untreated control cells.

The half maximal inhibitory concentration (IC ₅₀ ) of each oligonucleotide is also presented in the table below. ANGPTL3 mRNA levels were significantly decreased in a dose-dependent manner in antisense oligonucleotide-treated cells.

Table 156

Table 157

Table 158

Table 159

Example 123: Dose-dependent antisense inhibition of human ANGPTL3 in Hep3B cells

Deoxy, MOE, and cEt oligonucleotides were selected from the studies described above showing significant in vitro inhibition of ANGPTL3 mRNA and tested at various doses in Hep3B cells. ISIS 337847, 5-10-5 MOE gapmers were also included in this study. The antisense oligonucleotides were tested for potency in a series of parallel experiments with similar culture conditions. The results of each experiment are shown in a separate table below.

Cells were plated at a density of 20,000 cells per well and transfected using electroporation with antisense oligonucleotides at concentrations of 0.160 μM, 0.481 μM, 1.444 μM, 4.333 μM and 13.00 μM, as indicated in the table below. After a treatment period of approximately 16 hours, RNA was isolated from the cells and ANGPTL3 mRNA levels were measured by quantitative real-time PCR. mRNA levels were measured using the human primer probe set RTS3492_MGB. ANGPTL3 mRNA levels were adjusted according to total RNA content as measured by RiboGreen®. Results are expressed as percent inhibition of ANGPTL3 relative to untreated control cells.

The half maximal inhibitory concentration (IC ₅₀ ) of each oligonucleotide is also presented in the table below. ANGPTL3 mRNA levels were significantly decreased in a dose-dependent manner in antisense oligonucleotide-treated cells.

Table 160

Table 161

Table 162

Example 124: Dose-dependent antisense inhibition of human ANGPTL3 in Hep3B cells by MOE gapmers

MOE gapmers from the studies described above that showed significant in vitro inhibition of ANGPTL3 mRNA were selected and tested at various doses in Hep3B cells. Cells were plated at a density of 20,000 cells per well and transfected using electroporation with antisense oligonucleotides at concentrations of 0.160 μM, 0.481 μM, 1.444 μM, 4.333 μM and 13.00 μM, as indicated in the table below. After a treatment period of approximately 16 hours, RNA was isolated from the cells and ANGPTL3 mRNA levels were measured by quantitative real-time PCR. mRNA levels were measured using the human primer probe set RTS3492_MGB. ANGPTL3 mRNA levels were adjusted according to total RNA content as measured by ^{RiboGreen ®.} Results are expressed as percent inhibition of ANGPTL3 relative to untreated control cells.

The half maximal inhibitory concentration (IC ₅₀ ) of each oligonucleotide is also presented in the table below. ANGPTL3 mRNA levels were significantly decreased in a dose-dependent manner in antisense oligonucleotide-treated cells.

Table 163

Example 125: Dose-dependent antisense inhibition of human ANGPTL3 in Hep3B cells by deoxy, MOE and cEt oligonucleotides

Deoxy, MOE, and cEt oligonucleotides were selected from the studies described above showing significant in vitro inhibition of ANGPTL3 mRNA and tested at various doses in Hep3B cells. Cells were plated at a density of 20,000 cells per well and transfected using electroporation with antisense oligonucleotides at concentrations of 0.111 μM, 0.333 μM, 1.00 μM, 3.00 μM and 9.00 μM, as indicated in the table below. After a treatment period of approximately 16 hours, RNA was isolated from the cells and ANGPTL3 mRNA levels were measured by quantitative real-time PCR. mRNA levels were measured using the human primer probe set RTS3492_MGB. ANGPTL3 mRNA levels were adjusted according to total RNA content as measured by ^{RiboGreen ®.} Results are expressed as percent inhibition of ANGPTL3 relative to untreated control cells.

The half maximal inhibitory concentration (IC ₅₀ ) of each oligonucleotide is also presented in the table below. ANGPTL3 mRNA levels were significantly decreased in a dose-dependent manner in antisense oligonucleotide-treated cells.

Table 164

Table 165

Table 166

Table 167

Example 126: Antisense inhibition of human ANGPTL3 in huANGPTL3 transgenic mice

The antisense oligonucleotides described in this study were further evaluated for their ability to reduce the human ANGPTL3 mRNA transcript in C57Bl/6 mice carrying the human ANGPTL3 transgene (Tg mice).

Study 1

Female Tg mice were maintained on a 12-hour light/dark cycle. Animals were acclimatized for at least 7 days in the study facility prior to initiation of the experiment. Antisense oligonucleotides (ASO) were prepared in buffered saline (PBS) and sterilized by filtration through a 0.2 micron filter. Oligonucleotides were dissolved in 0.9% PBS for injection.

Groups of mice were given an intraperitoneal injection of 5-10-5 MOE gapmer once per week at a dose of 50 mg/kg for 2 weeks. One group of mice received a subcutaneous injection of PBS once per week for 2 weeks. The PBS-injected group served as a control group to compare the corresponding oligonucleotide-treated groups.

RNA analysis

At the end of the treatment period, RNA was extracted from the liver for real-time PCR analysis of the measurement of mRNA expression of ANGPTL3 with RTS3492_MGB. mRNA levels were also determined by the human primer probe set RTS1984 (forward sequence CTTCAATGAAACGTGGGAACT, designated herein as SEQ ID NO: 7; reverse sequence TCTCTAGGCCCAACCAAAATTC, designated herein as SEQ ID NO: 8; probe sequence AAATATGGTTTTGGGAGGCTTGAT, designated herein as SEQ ID NO: 9 ) was measured. Results are expressed as percent change in mRNA compared to PBS control and normalized to ^{RiboGreen ®.} As shown in the table below, treatment with ISIS antisense oligonucleotides resulted in a significant reduction in ANGPTL3 mRNA compared to the PBS control.

Table 168

% inhibition of ANGPTL3 mRNA in the liver of transgenic mice compared to the PBS control group

protein analysis

Human ANGPTL3 protein levels were quantified using a commercially available ELISA kit (product number #DANL30 by R&D Systems, Minneapolis, Minn.) with transformed plasma samples diluted 1:20,000 using the protocol described by the manufacturer. The results are shown in the table below. These results indicate that treatment with ISIS oligonucleotides resulted in reduced ANGPTL3 protein levels.

Table 169

% Inhibition of Plasma Protein Levels in Transgenic Mice

Plasma Chemistry Markers

To evaluate the effects of ISIS oligonucleotides on day 10, plasma levels of transaminases (ALT and AST) were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS oligonucleotides that caused any change in the level of any of these liver function markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 170

Plasma transaminase levels (IU/L) in transgenic mice at day 10

study 2

Male Tg mice were maintained on a 12-hour light/dark cycle. Animals were acclimatized for at least 7 days in the study facility prior to initiation of the experiment. Antisense oligonucleotides (ASO) were prepared in buffered saline (PBS) and sterilized by filtration through a 0.2 micron filter. Oligonucleotides were dissolved in 0.9% PBS for injection.

Groups of mice were given an intraperitoneal injection of 5-10-5 MOE gapmer once per week at a dose of 50 mg/kg for 2 weeks. One group of mice received a subcutaneous injection of PBS once per week for 2 weeks. The PBS-injected group served as a control group to compare the corresponding oligonucleotide-treated groups.

RNA analysis

At the end of the treatment period, RNA was extracted from the liver for real-time PCR analysis of the measurement of mRNA expression of ANGPTL3 with RTS1984. Results are expressed as percent change in mRNA compared to PBS control and normalized to ^{RiboGreen ®.} As shown in the table below, treatment with ISIS antisense oligonucleotides resulted in a significant reduction in ANGPTL3 mRNA compared to the PBS control.

Table 171

% inhibition of ANGPTL3 mRNA in the liver of transgenic mice compared to the PBS control group

protein analysis

Human ANGPTL3 protein levels were quantified using a commercially available ELISA kit (product number #DANL30 by R&D Systems, Minneapolis, Minn.) with transformed plasma samples diluted 1:20,000 using the protocol described by the manufacturer. The results are shown in the table below. These results indicate that treatment with ISIS oligonucleotides resulted in reduced ANGPTL3 protein levels.

Table 172

% Inhibition of Plasma Protein Levels in Transgenic Mice

Plasma Chemistry Markers

To evaluate the effects of ISIS oligonucleotides on day 8, plasma levels of transaminases (ALT and AST) were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS oligonucleotides that caused any change in the level of any of these liver function markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 173

Plasma transaminase levels (IU/L) in transgenic mice at day 8

Study 3

Male and female Tg mice were maintained on a 12 hour light/dark cycle. Animals were acclimatized for at least 7 days in the study facility prior to initiation of the experiment. Antisense oligonucleotides (ASO) were prepared in buffered saline (PBS) and sterilized by filtration through a 0.2 micron filter. Oligonucleotides were dissolved in 0.9% PBS for injection.

Groups of mice were given intraperitoneal injections of 5-10-5 MOE gapmers at doses of 2.5 mg/kg, 12.5 mg/kg, or 25 mg/kg once per week for 3 weeks. One group of mice received a subcutaneous injection of PBS once per week for 2 weeks. The PBS-injected group served as a control group to compare the corresponding oligonucleotide-treated groups.

RNA analysis

At the end of the treatment period, RNA was extracted from the liver for real-time PCR analysis of measurement of mRNA expression of ANGPTL3 with hANGPTL3_LTS01022 (forward sequence AAATTTTAGCCAATGGCCTCC, designated herein as SEQ ID NO: 10; TGTCATTAATTTGGCCCTTCG, reverse sequence, with SEQ ID NO: 11) Designated herein; probe sequence TCAGTTGGGACATGGTCTTAAAGACTTTGTCC, designated SEQ ID NO: 12). Results are expressed as percent change in mRNA compared to PBS control and normalized to ^{RiboGreen ®.} As shown in the table below, treatment with ISIS antisense oligonucleotides resulted in a significant reduction in ANGPTL3 mRNA compared to the PBS control. The ED ₅₀ of each gapmer is also shown in the table below. 'nd' indicates that the ED ₅₀ was not measured.

Table 174

% inhibition of ANGPTL3 mRNA in the liver of transgenic mice compared to the PBS control group

protein analysis

Human ANGPTL3 protein levels were quantified using a commercially available ELISA kit (product number #DANL30 by R&D Systems, Minneapolis, Minn.) with transformed plasma samples diluted 1:20,000 using the protocol described by the manufacturer. The results are shown in the table below. These results indicate that treatment with ISIS oligonucleotides resulted in reduced ANGPTL3 protein levels. 'nd' indicates that no _{ED 50 was measured.}

Table 175

% Inhibition of Plasma Protein Levels in Transgenic Mice

Plasma Chemistry Markers

To evaluate the effects of ISIS oligonucleotides on day 17, plasma levels of transaminases (ALT and AST) were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS oligonucleotides that caused any change in the level of any of these liver function markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 176

Plasma transaminase levels (IU/L) in transgenic mice at day 17

study 4

Male and female Tg mice were maintained on a 12 hour light/dark cycle. Animals were acclimatized for at least 7 days in the study facility prior to initiation of the experiment. Antisense oligonucleotides (ASO) were prepared in buffered saline (PBS) and sterilized by filtration through a 0.2 micron filter. Oligonucleotides were dissolved in 0.9% PBS for injection.

Groups of mice were given intraperitoneal injections of 5-10-5 MOE gapmers at a dose of 25 mg/kg once per week for 2 weeks. One group of mice received a subcutaneous injection of PBS once per week for 2 weeks. The PBS-injected group served as a control group to compare the corresponding oligonucleotide-treated groups.

RNA analysis

At the end of the treatment period, RNA was extracted from the liver for real-time PCR analysis of the measurement of mRNA expression of ANGPTL3 with hANGPTL3_LTS01022. Results are expressed as percent change in mRNA compared to PBS control and normalized to ^{RiboGreen ®.} As shown in the table below, treatment with ISIS antisense oligonucleotides resulted in a significant reduction in ANGPTL3 mRNA compared to the PBS control.

Table 177

% inhibition of ANGPTL3 mRNA in the liver of transgenic mice compared to the PBS control group

Plasma Chemistry Markers

To evaluate the effects of ISIS oligonucleotides on day 10, plasma levels of transaminases (ALT and AST) were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS oligonucleotides that caused any change in the level of any of these liver function markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 178

Plasma transaminase levels (IU/L) in transgenic mice at day 10

study 5

Male and female Tg mice were maintained on a 12 hour light/dark cycle. Animals were acclimatized for at least 7 days in the study facility prior to initiation of the experiment. Antisense oligonucleotides (ASO) were prepared in buffered saline (PBS) and sterilized by filtration through a 0.2 micron filter. Oligonucleotides were dissolved in 0.9% PBS for injection.

Groups of mice received intraperitoneal injections of 5-10-5 MOE gapmers or deoxy, MOE, and cEt gapmers at a dose of 25 mg/kg once per week for 2 weeks. One group of mice received a subcutaneous injection of PBS once per week for 2 weeks. The PBS-injected group served as a control group to compare the corresponding oligonucleotide-treated groups.

RNA analysis

At the end of the treatment period, RNA was extracted from the liver for real-time PCR analysis of the measurement of mRNA expression of ANGPTL3 with RTS1984. Results are expressed as percent change in mRNA compared to PBS control and normalized to ^{RiboGreen ®.} As shown in the table below, treatment with ISIS antisense oligonucleotides resulted in a significant reduction in ANGPTL3 mRNA compared to the PBS control.

Table 179

% inhibition of ANGPTL3 mRNA in the liver of transgenic mice compared to the PBS control group

Plasma Chemistry Markers

To evaluate the effects of ISIS oligonucleotides on day 9, plasma levels of transaminases (ALT and AST) were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS oligonucleotides that caused any change in the level of any of these liver function markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 180

Plasma transaminase levels (IU/L) in transgenic mice at day 9

study 6

Male and female Tg mice were maintained on a 12 hour light/dark cycle. Animals were acclimatized for at least 7 days in the study facility prior to initiation of the experiment. Antisense oligonucleotides (ASO) were prepared in buffered saline (PBS) and sterilized by filtration through a 0.2 micron filter. Oligonucleotides were dissolved in 0.9% PBS for injection.

Groups of mice were given intraperitoneal injections of deoxy, MOE, and cEt oligonucleotides at a dose of 25 mg/kg once per week for 2 weeks. ISIS 233710, 5-10-5 MOE gapmers were also included as benchmarks. One group of mice received a subcutaneous injection of PBS once per week for 2 weeks. The PBS-injected group served as a control group to compare the corresponding oligonucleotide-treated groups.

RNA analysis

At the end of the treatment period, RNA was extracted from the liver for real-time PCR analysis of the measurement of mRNA expression of ANGPTL3 with hANGPTL3_LTS01022. Results are expressed as percent change in mRNA compared to PBS control and normalized to ^{RiboGreen ®.} As shown in the table below, treatment with several ISIS antisense oligonucleotides resulted in a significant decrease in ANGPTL3 mRNA compared to the PBS control.

Table 181

% inhibition of ANGPTL3 mRNA in the liver of transgenic mice compared to the PBS control group

protein analysis

Human ANGPTL3 protein levels were quantified using a commercially available ELISA kit (product number #DANL30 by R&D Systems, Minneapolis, Minn.) with transformed plasma samples diluted 1:20,000 using the protocol described by the manufacturer. The results are shown in the table below. These results indicate that treatment with several ISIS oligonucleotides resulted in reduced ANGPTL3 protein levels.

Table 182

% Inhibition of Plasma Protein Levels in Transgenic Mice

Plasma Chemistry Markers

To evaluate the effects of ISIS oligonucleotides on day 10, plasma levels of transaminases (ALT and AST) were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS oligonucleotides that caused any change in the level of any of these liver function markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 183

Plasma transaminase levels (IU/L) in transgenic mice at day 10

study 7

Male and female Tg mice were maintained on a 12 hour light/dark cycle. Animals were acclimatized for at least 7 days in the study facility prior to initiation of the experiment. Antisense oligonucleotides (ASO) were prepared in buffered saline (PBS) and sterilized by filtration through a 0.2 micron filter. Oligonucleotides were dissolved in 0.9% PBS for injection.

Groups of mice were given intraperitoneal injections of deoxy, MOE, and cEt oligonucleotides at a dose of 25 mg/kg once per week for 2 weeks. ISIS 233710, 5-10-5 MOE gapmers were also included as benchmarks. One group of mice received a subcutaneous injection of PBS once per week for 2 weeks. The PBS-injected group served as a control group to compare the corresponding oligonucleotide-treated groups.

RNA analysis

At the end of the treatment period, RNA was extracted from the liver for real-time PCR analysis of the measurement of mRNA expression of ANGPTL3 with hANGPTL3_LTS01022. Results are expressed as percent change in mRNA compared to PBS control and normalized to ^{RiboGreen ®.} As shown in the table below, treatment with ISIS antisense oligonucleotides resulted in a significant reduction in ANGPTL3 mRNA compared to the PBS control.

Table 184

% inhibition of ANGPTL3 mRNA in the liver of transgenic mice compared to the PBS control group

protein analysis

Human ANGPTL3 protein levels were quantified using a commercially available ELISA kit (product number #DANL30 by R&D Systems, Minneapolis, Minn.) with transformed plasma samples diluted 1:20,000 using the protocol described by the manufacturer. The results are shown in the table below. These results indicate that treatment with ISIS oligonucleotides resulted in reduced ANGPTL3 protein levels. In this case, a value of '0' indicates that treatment with some of the ISIS oligonucleotides did not inhibit expression; In some instances, increased levels of expression can be recorded.

Table 185

% Inhibition of Plasma Protein Levels in Transgenic Mice

Plasma Chemistry Markers

To evaluate the effects of ISIS oligonucleotides on day 9, plasma levels of transaminases (ALT and AST) were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS oligonucleotides that caused any change in the level of any of these liver function markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 186

Plasma transaminase levels (IU/L) in transgenic mice at day 9

study 8

Male and female Tg mice were maintained on a 12 hour light/dark cycle. Animals were acclimatized for at least 7 days in the study facility prior to initiation of the experiment. Antisense oligonucleotides (ASO) were prepared in buffered saline (PBS) and sterilized by filtration through a 0.2 micron filter. Oligonucleotides were dissolved in 0.9% PBS for injection.

Groups of mice were given intraperitoneal injections of deoxy, MOE, and cEt oligonucleotides at a dose of 25 mg/kg once per week for 2 weeks. ISIS 233710, 5-10-5 MOE gapmers were also included as benchmarks. One group of mice received a subcutaneous injection of PBS once per week for 2 weeks. The PBS-injected group served as a control group to compare the corresponding oligonucleotide-treated groups.

RNA analysis

At the end of the treatment period, RNA was extracted from the liver for real-time PCR analysis of the measurement of mRNA expression of ANGPTL3 with hANGPTL3_LTS01022. Results are expressed as percent change in mRNA compared to PBS control and normalized to ^{RiboGreen ®.} As shown in the table below, treatment with ISIS antisense oligonucleotides resulted in a significant reduction in ANGPTL3 mRNA compared to the PBS control.

Table 187

% inhibition of ANGPTL3 mRNA in the liver of transgenic mice compared to the PBS control group

protein analysis

Human ANGPTL3 protein levels were quantified using a commercially available ELISA kit (product number #DANL30 by R&D Systems, Minneapolis, Minn.) with transformed plasma samples diluted 1:20,000 using the protocol described by the manufacturer. The results are shown in the table below. These results indicate that treatment with ISIS oligonucleotides produced reduced ANGPTL3 levels.

Table 188

% inhibition of plasma protein levels in transgenic mice

Plasma Chemistry Markers

To evaluate the effects of ISIS oligonucleotides on day 8, plasma levels of transaminases (ALT and AST) were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS oligonucleotides that caused any change in the level of any of these liver function markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 189

Plasma transaminase levels (IU/L) in transgenic mice at day 8

study 9

Male and female Tg mice were maintained on a 12 hour light/dark cycle. Animals were acclimatized for at least 7 days in the study facility prior to initiation of the experiment. Antisense oligonucleotides (ASO) were prepared in buffered saline (PBS) and sterilized by filtration through a 0.2 micron filter. Oligonucleotides were dissolved in 0.9% PBS for injection.

Groups of mice were given intraperitoneal injections of deoxy, MOE, and cEt oligonucleotides at a dose of 25 mg/kg once per week for 2 weeks. ISIS 233710, 5-10-5 MOE gapmers were also included as benchmarks. One group of mice received a subcutaneous injection of PBS once per week for 2 weeks. The PBS-injected group served as a control group to compare the corresponding oligonucleotide-treated groups.

RNA analysis

At the end of the treatment period, RNA was extracted from the liver for real-time PCR analysis of the measurement of mRNA expression of ANGPTL3 with hANGPTL3_LTS01022. Results are expressed as percent change in mRNA compared to PBS control and normalized to ^{RiboGreen ®.} As shown in the table below, treatment with ISIS antisense oligonucleotides resulted in a significant decrease in ANGPTL3 mRNA compared to the PBS control. In this case, a value of '0' indicates that treatment with some of the ISIS oligonucleotides did not inhibit expression; In some instances, increased levels of expression can be recorded.

Table 190

% inhibition of ANGPTL3 mRNA in the liver of transgenic mice compared to the PBS control group

protein analysis

Human ANGPTL3 protein levels were quantified using a commercially available ELISA kit (product number #DANL30 by R&D Systems, Minneapolis, Minn.) with transformed plasma samples diluted 1:20,000 using the protocol described by the manufacturer. The results are shown in the table below. These results indicate that treatment with ISIS oligonucleotides resulted in reduced ANGPTL3 protein levels. In this case, a value of '0' indicates that treatment with some of the ISIS oligonucleotides did not inhibit expression; In some instances, increased levels of expression can be recorded.

Table 191

% inhibition of plasma protein levels in transgenic mice

Plasma Chemistry Markers

To evaluate the effects of ISIS oligonucleotides on day 9, plasma levels of transaminases (ALT and AST) were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS oligonucleotides that caused any change in the level of any of these liver function markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 192

Plasma transaminase levels (IU/L) in transgenic mice at day 9

study 10

Male and female Tg mice were maintained on a 12 hour light/dark cycle. Animals were acclimatized for at least 7 days in the study facility prior to initiation of the experiment. Antisense oligonucleotides (ASO) were prepared in buffered saline (PBS) and sterilized by filtration through a 0.2 micron filter. Oligonucleotides were dissolved in 0.9% PBS for injection.

Groups of mice were given intraperitoneal injections of 5-10-5 MOE gapmer or deoxy, MOE, and cEt oligonucleotides at doses of 5 mg/kg, 12.5 mg/kg, or 25 mg/kg once per week for 2 weeks. provided. One group of mice received a subcutaneous injection of PBS once per week for 2 weeks. The PBS-injected group served as a control group to compare the corresponding oligonucleotide-treated groups.

RNA analysis

At the end of the treatment period, RNA was extracted from the liver for real-time PCR analysis of the measurement of mRNA expression of ANGPTL3 with hANGPTL3_LTS01022 and RTS3492_MGB as well. Results are expressed as percent change in mRNA compared to PBS control and normalized to ^{RiboGreen ®.} As shown in the table below, treatment with some of the ISIS antisense oligonucleotides resulted in a significant reduction in ANGPTL3 mRNA compared to the PBS control.

Table 193

% inhibition of ANGPTL3 mRNA in the liver of transgenic mice compared to the PBS control group

Plasma Chemistry Markers

To evaluate the effects of ISIS oligonucleotides on day 8, plasma levels of transaminases (ALT and AST) were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS oligonucleotides that caused any change in the level of any of these liver function markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 194

Plasma transaminase levels (IU/L) in transgenic mice at day 8

Example 127: Resistance of antisense oligonucleotides targeting human ANGPTL3 in CD1 mice

The CD1® mouse (Charles River, MA) is a multipurpose mouse model and is often used for safety and efficacy testing. The mice were treated with ISIS antisense oligonucleotides selected from the studies described above and changes in levels of various plasma chemical markers were assessed.

Study 1

Male CD1 mice (one animal per treatment group) were injected intraperitoneally with deoxy, MOE, and cEt oligonucleotides at a single dose of 200 mg/kg. One male CD1 mouse was injected subcutaneously with a single dose of PBS. Mice were euthanized 48 hours after the last dose and organs and plasma were harvested for further analysis.

Plasma Chemistry Markers

To evaluate the effects of ISIS oligonucleotides on day 4, plasma levels of transaminases (ALT and AST) were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS oligonucleotides that caused any change in the level of any of these liver function markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 195

Plasma transaminase levels in CD1 mice at day 4

weight

Body weights were measured on day 1 after the daily dose of ISIS oligonucleotides and are shown in the table below. ISIS oligonucleotides that caused certain organ weight changes outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 196

Body weight of CD1 mice after antisense oligonucleotide treatment (g)

study 2

Male CD1 mice (one animal per treatment group) were injected intraperitoneally with deoxy, MOE and cEt oligonucleotides at a single dose of 200 mg/kg. One male CD1 mouse was injected subcutaneously with a single dose of PBS. Mice were euthanized 48 hours after the last dose and organs and plasma were harvested for further analysis.

Plasma Chemistry Markers

To evaluate the effects of ISIS oligonucleotides on day 5, plasma levels of transaminases (ALT and AST) were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS oligonucleotides that caused any change in the level of any of these liver function markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 197

Plasma transaminase levels in CD1 mice at day 5

weight

Body weight was measured on day 5 after the daily dose of ISIS oligonucleotide and is shown in the table below. ISIS oligonucleotides that caused certain organ weight changes outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 198

Body weight of CD1 mice after antisense oligonucleotide treatment (g)

study 3

Male CD1 mice (4 animals per treatment group) were injected intraperitoneally with 5-10-5 MOE gapmer at 100 mg/kg once per week for 6 weeks. One group of 4 male CD1 mice was injected intraperitoneally with PBS once per week for 6 weeks. Mice were euthanized 48 hours after the last dose and organs and plasma were harvested for further analysis.

Plasma Chemistry Markers

To evaluate the efficacy of ISIS oligonucleotides, plasma levels of various hepatic and renal function markers were measured at day 45 using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS oligonucleotides that caused any change in the level of any of these markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 199

Plasma Chemistry Marker Levels in CD1 Mice at Day 45

weight

The body weight was measured on day 43 and shown in the table below. Kidney, liver and spleen weights were measured at the end of the study on day 45. ISIS oligonucleotides that caused certain organ weight changes outside the expected range for antisense oligonucleotides were excluded from further studies.

table 200

Weight (g) of CD1 mice after antisense oligonucleotide treatment

study 4

Male CD1 mice (4 animals per treatment group) were injected intraperitoneally with 50 mg/kg or 100 mg/kg of 5-10-5 MOE gapmer or deoxy, MOE and cEt oligonucleotides given once a week for 6 weeks. One group of 4 male CD1 mice was injected intraperitoneally with PBS once per week for 6 weeks. Mice were euthanized 48 hours after the last dose and organs and plasma were harvested for further analysis.

Plasma Chemistry Markers

To evaluate the efficacy of ISIS oligonucleotides, plasma levels of various hepatic and renal function markers were measured at day 46 using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS oligonucleotides that caused any change in the level of any of these markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 201

Plasma Chemistry Marker Levels in CD1 Mice at Day 45

weight

The body weight was measured on day 44 and shown in the table below. Kidney, liver and spleen weights were measured at the end of the study on day 46. ISIS oligonucleotides that caused certain organ weight changes outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 202

Weight (g) of CD1 mice after antisense oligonucleotide treatment

study 5

Male CD1 mice (4 animals per treatment group) were injected intraperitoneally with 50 mg/kg of deoxy, MOE and cEt oligonucleotides given once a week for 6 weeks. One group of 4 male CD1 mice was injected intraperitoneally with PBS once per week for 6 weeks. Mice were euthanized 48 hours after the last dose and organs and plasma were harvested for further analysis.

Plasma Chemistry Markers

To evaluate the efficacy of ISIS oligonucleotides, plasma levels of various hepatic and renal function markers were measured on day 43 using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS oligonucleotides that caused any change in the level of any of these markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 203

Plasma Chemistry Marker Levels in CD1 Mice at Day 43

weight

The body weight was measured on day 41 and shown in the table below. Kidney, liver and spleen weights were measured at the end of the study on day 43. ISIS oligonucleotides that caused certain organ weight changes outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 204

Weight (g) of CD1 mice after antisense oligonucleotide treatment

Example 128: Determination of Viscosity of ISIS Antisense Oligonucleotides Targeting Human ANGPTL3

The viscosity of selected antisense oligonucleotides from the studies described above was measured for the purpose of screening antisense oligonucleotides having a centipoise (cP) of 40 or greater. Oligonucleotides with a viscosity greater than 40 cP will have a less than optimal viscosity.

ISIS oligonucleotides into a glass vial to improve the nucleotide (32-35 mg), was dissolved in an antisense oligo nucleotide into solution by the addition of 120 μL of water and heating the vial at 50 ⁰ C. A portion (75 μL) of the preheated sample was pipetted into a micro-viscometer (Cambridge). Micro-set the temperature of the viscometer to 25 ⁰ C, which was the viscosity of the sample. Another portion (20 μL) of pre-heated sample was pipetted into 10 mL of water for UV reading at 260 nM at ^{85 0 C (Cary UV device).} The results are presented in the table below, where the concentration of each antisense oligonucleotide was 350 mg/ml, indicating that most of the antisense oligonucleotide solutions are optimal for their viscosity within the criteria described above.

Table 205

Viscosity of ISIS antisense oligonucleotides targeting human ANGPTL3

Example 129: Resistance of antisense oligonucleotides targeting human ANGPTL3 in Sprague-Dawley rats

The Sprague-Dawley rat is a versatile model used for safety and efficacy evaluation. Rats were treated with ISIS antisense oligonucleotides from the studies described in the Examples above and changes in the levels of various plasma chemical markers were assessed.

Study 1

Male Sprague-Dawley rats were maintained on a 12-hour light/dark cycle and fed ad libitum Purina normal rat chow, Diet 5001. Groups of 4 Sprague-Dawley rats each were injected subcutaneously with PBS or 100 mg/kg of 5-10-5 MOE gapmer once per week for 6 weeks. At 48 hours after the last dose, the rats were euthanized and plasma was collected for further analysis.

liver function

To evaluate the efficacy of ISIS oligonucleotides on liver function, plasma levels of transaminase were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). Plasma levels of ALT (alanine transaminase) and AST (aspartase transaminase) were measured on day 45 and the results are presented in IU/L in the table below. Plasma levels of bilirubin were also measured using the same clinical chemistry analyzer and the results are also presented in mg/dL in the table below. ISIS oligonucleotides that caused any change in any level of any of the liver function markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 206

Hepatic function markers in Sprague-Dawley rats

kidney function

To evaluate the effect of ISIS oligonucleotides on renal function, plasma levels of blood urea nitrogen (BUN) and creatinine were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in mg/dL in the table below. ISIS oligonucleotides that caused any change in any level of any of the renal function markers outside the expected range for antisense oligonucleotides were excluded from further studies. Total urine protein and urine creatinine levels were measured, and the ratio of total urine protein to creatinine was evaluated. The results are shown in the table below.

Table 207

Renal Function Plasma Markers in Sprague-Dawley Rats (mg/dL)

Table 208

Renal Function Urine Markers in Sprague-Dawley Rats

long-term weight

Body weight was measured on day 42 and shown in the table below. Liver, spleen and kidney weights were measured at the end of the study on day 45 and are presented in the table below. ISIS oligonucleotides that caused certain organ weight changes outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 209

Body and Organ Weights of Sprague-Dawley Rats (g)

study 2

Male Sprague-Dawley rats were maintained on a 12-hour light/dark cycle and fed ad libitum Purina normal rat chow, Diet 5001. Groups of 4 Sprague-Dawley rats each were injected subcutaneously with PBS or 50 mg/kg or 100 mg/kg of 5-10-5 MOE gapmer or deoxy, MOE and cEt oligonucleotides once per week for 6 weeks. At 48 hours after the last dose, the rats were euthanized and plasma was collected for further analysis.

liver function

To evaluate the efficacy of ISIS oligonucleotides on liver function, plasma levels of transaminase were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). Plasma levels of ALT (alanine transaminase) and AST (aspartase transaminase) were measured on day 44 and the results are presented in IU/L in the table below. Plasma levels of bilirubin were also measured using the same clinical chemistry analyzer and the results are also presented in mg/dL in the table below. ISIS oligonucleotides that caused any change in any level of any of the liver function markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 210

Hepatic function markers in Sprague-Dawley rats

kidney function

To evaluate the effect of ISIS oligonucleotides on renal function, plasma levels of blood urea nitrogen (BUN) and creatinine at day 44 were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). . The results are shown in mg/dL in the table below. ISIS oligonucleotides that caused any change in any level of any of the renal function markers outside the expected range for antisense oligonucleotides were excluded from further studies. Total urine protein and urine creatinine levels were measured, and the ratio of total urine protein to creatinine was evaluated. The results are shown in the table below.

Table 211

Renal Function Plasma Markers in Sprague-Dawley Rats (mg/dL)

Table 212

Renal Function Urine Markers in Sprague-Dawley Rats

long-term weight

Body weight was measured on day 42 and shown in the table below. Liver, spleen and kidney weights were measured at the end of the study on day 44 and are presented in the table below. ISIS oligonucleotides that caused certain organ weight changes outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 213

Body and Organ Weights of Sprague-Dawley Rats (g)

Study 3

Male Sprague-Dawley rats were maintained on a 12-hour light/dark cycle and fed ad libitum Purina normal rat chow, Diet 5001. Groups of 4 Sprague-Dawley rats each were injected subcutaneously with PBS or 50 mg/kg of deoxy, MOE and cEt oligonucleotides once per week for 6 weeks. At 48 hours after the last dose, the rats were euthanized and plasma was collected for further analysis.

liver function

To evaluate the efficacy of ISIS oligonucleotides on liver function, plasma levels of transaminase were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). Plasma levels of ALT (alanine transaminase) and AST (aspartase transaminase) were measured on day 44 and the results are presented in IU/L in the table below. Plasma levels of bilirubin were also measured using the same clinical chemistry analyzer and the results are also presented in mg/dL in the table below. ISIS oligonucleotides that caused any change in any level of any of the liver function markers outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 214

Hepatic function markers in Sprague-Dawley rats

kidney function

To evaluate the effect of ISIS oligonucleotides on renal function, plasma levels of blood urea nitrogen (BUN) and creatinine at day 44 were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). . The results are shown in mg/dL in the table below. ISIS oligonucleotides that caused any change in any level of any of the renal function markers outside the expected range for antisense oligonucleotides were excluded from further studies. Total urine protein and urine creatinine levels were measured, and the ratio of total urine protein to creatinine was evaluated. The results are shown in the table below.

Table 215

Renal Function Plasma Markers in Sprague-Dawley Rats (mg/dL)

Table 216

Renal Function Urine Markers in Sprague-Dawley Rats

long-term weight

Body weight was measured on day 42 and shown in the table below. Liver, spleen and kidney weights were measured at the end of the study on day 44 and are presented in the table below. ISIS oligonucleotides that caused certain organ weight changes outside the expected range for antisense oligonucleotides were excluded from further studies.

Table 217

Body and Organ Weights of Sprague-Dawley Rats (g)

Example 130: Effect of ISIS antisense oligonucleotides targeting human ANGPTL3 in cynomolgus monkeys

Cynomolgus monkeys were treated with ISIS antisense oligonucleotides selected from the studies described in the Examples above. Antisense oligonucleotide efficacy and tolerability, and also their pharmacodynamic profile in liver and kidney were evaluated.

At the time this study was undertaken, cynomolgus monkey genome sequences were not available in the National Center for Biotechnology Information (NCBI) database, so cross-reactivity with cynomolgus monkey gene sequences could not be confirmed. Instead, the sequence of the ISIS antisense oligonucleotide used in cynomolgus monkeys was compared to the rhesus sequence for homology. ISIS oligonucleotides homologous to rhesus sequences are also expected to fully cross-react with cynomolgus monkey sequences. The human antisense oligonucleotides tested are cross-reactive with a rhesus genomic sequence (GenBank Accession No. NW_001108682.1, truncated from nucleotides 3049001-3062000, designated herein as SEQ ID NO: 3). The greater the complementarity between the human oligonucleotide and the rhesus sequence, the more likely the human oligonucleotide can cross-react with the rhesus sequence. The start and stop sites of each oligonucleotide for SEQ ID NO: 3 are shown in the table below. "Initiation site" refers to the 5'-maximum nucleotide to which the gapmer is targeted within the rhesus monkey gene sequence. 'Mismatch' refers to the number of nucleobases in human oligonucleotides that mismatch the rhesus genomic sequence.

Table 218

Antisense oligonucleotide complementary to rhesus ANGPTL3 gene sequence (SEQ ID NO: 3)

cure

Prior to the study, the monkeys were kept in isolation for a period of at least 30 days, during which the animals were observed daily for general health. The monkeys were 2-4 years old and weighed between 2-4 kg. Nine groups of 5 randomly assigned male cynomolgus monkeys each received ISIS oligonucleotides or PBS at 4 sites (i.e., left, upper, right, and lower) in a clockwise direction, 1 site per dose. were injected subcutaneously. Monkeys were given a loading dose of PBS or 40 mg/kg of ISIS oligonucleotides every 2 days (days 1, 3, 5, and 7) for the first week and once per week for 12 weeks (14, 21, 28, 35). , 42, 49, 56, 63, 70, 77, and 84 days) PBS or 40 mg/kg of ISIS oligonucleotides were continuously administered.

During the study period, monkeys were observed twice daily for disease or distress. Certain animals experiencing transient or mild pain or distress abnormalities due to treatment, injury or illness were treated with an analgesic or formulation approved by the veterinary staff for pain relief after consultation with the study leader. Certain animals in unhealthy or possible moribund conditions were confirmed for further observation and possible euthanasia. For example, one animal in the ISIS 567321 treatment group was found to be moribund on day 45 and terminated. Scheduled euthanasia of animals was performed on day 86 (approximately 48 hours after the last dose) by ketamine/xylazine-induced anesthesia and exsanguination following administration of sodium pentobarbital. The protocol described in this example was approved by the Animal Experimental Ethics Committee (IACUC).

reduced liver target

RNA analysis

On day 86, RNA was extracted from liver for real-time PCR analysis of the measurement of mRNA expression of ANGPTL3. Results are expressed as percent change in mRNA compared to PBS control and normalized to ^{RiboGreen ®.} As shown in the table below, treatment with ISIS antisense oligonucleotides resulted in a significant reduction in ANGPTL3 mRNA compared to the PBS control. Analysis of ANGPTL3 mRNA levels indicates that ISIS 544199 and ISIS 559277, both fully cross-reactive with the rhesus sequence, significantly reduced expression levels. Other ISIS oligonucleotides that target mismatched monkey sequences may also decrease ANGPTL3 mRNA levels.

Table 219

% inhibition of ANGPTL3 mRNA in cynomolgus monkey liver compared to PBS control

protein analysis

Approximately 1 mL of blood was collected from all available animals on day 85 and placed in tubes containing the potassium salt of EDTA. Plasma was collected by placing the blood sample on ice and centrifuging (3000 rpm for 10 min at 4°C).

Human ANGPTL3 protein levels were quantified using a commercially available ELISA kit (product number #DANL30 by R&D Systems, Minneapolis, Minn.) with transformed plasma samples diluted 1:20,000 using the protocol described by the manufacturer. The results are shown in the table below. Analysis of plasma ANGPTL3 showed that ISIS 563580, 544199 and ISIS 559277 reduced protein levels in a delayed manner. Other ISIS oligonucleotides were also able to reduce ANGPTL3 levels.

Table 220

Plasma protein levels in cynomolgus monkeys (ng/mL)

tolerance study

weight measurement

In order to evaluate the effect of ISIS oligonucleotides on the overall health of animals, the size and body weight were measured and presented in the table below. These results indicate that the effect of treatment with antisense oligonucleotides on body weight was within the predicted range for antisense oligonucleotides. Specifically, treatment with ISIS 563580 was also very tolerable in terms of monkey body weight.

Table 221

Final weight in cynomolgus monkeys (g)

liver function

To evaluate the effect of ISIS oligonucleotides on liver function, blood samples were collected from all study groups. Blood samples were collected 48 hours post-dose from the coccygeal, saphenous, or femoral veins. The monkeys were fasted overnight prior to blood collection. Blood was collected in tubes without anticoagulant for serum separation. Serum was collected by holding the tube at room temperature for a minimum of 90 minutes followed by centrifugation (at approximately 3,000 rpm for 10 minutes). The levels of various liver function markers were measured using a Toshiba 200FR NEO Chemistry analyzer (Toshiba Co., Japan). Plasma levels of ALT and AST were measured and the results are expressed in IU/L in the table below. A liver function marker, bilirubin, was similarly measured and expressed as mg/dL in the table below. These results indicate that most antisense oligonucleotides had no effect on liver function outside the range predicted for antisense oligonucleotides. Specifically, treatment with ISIS 563580 was highly resistant in terms of liver function in monkeys.

Table 222

ALT Levels in Cynomolgus Monkey Plasma (IU/L)

Table 223

AST Levels in Cynomolgus Monkey Plasma (IU/L)

Table 224

Cynomolgus monkey plasma bilirubin levels (mg/dL)

kidney function

To evaluate the effect of ISIS oligonucleotides on renal function, blood samples were collected from all study groups. Blood samples were collected 48 hours post-dose from the coccygeal, saphenous, or femoral veins. The monkeys were fasted overnight prior to blood collection. Blood was collected in tubes without anticoagulant for serum separation. Serum was collected by holding the tube at room temperature for a minimum of 90 minutes followed by centrifugation (at approximately 3,000 rpm for 10 minutes). The levels of BUN and creatinine were measured using a Toshiba 200FR NEO chemical analyzer (manufactured by Toshiba Co., Japan). The results are shown in mg/dL in the table below.

Plasma chemistry indicates that most of the ISIS oligonucleotides did not have any effect on renal function outside the predicted range for antisense oligonucleotides. Specifically, treatment with ISIS 563580 was highly resistant in terms of renal function in monkeys.

Table 225

Plasma BUN levels in cynomolgus monkeys (mg/dL)

Table 226

Plasma creatinine levels in cynomolgus monkeys (mg/dL)

hematology

To also evaluate any effect of ISIS oligonucleotides in cynomolgus monkeys on hematological parameters, blood samples of approximately 0.5 mL of blood were collected from each of the available study animals in tubes containing _{K 2 -EDTA.} Samples were analyzed for red blood cell (RBC) count, white blood cell (WBC) count, individual white blood cell count, such as that of monocytes, neutrophils, lymphocytes, and platelet count, hemoglobin content and hematocrit using an ADVIA120 Hematology Analyzer (Bayer, USA) analyzed. The data are shown in the table below.

The data indicate that the oligonucleotide did not cause any change in hematological parameters outside the range predicted for antisense oligonucleotides at this dose. Specifically, treatment with ISIS 563580 was highly resistant in terms of hematological parameters in monkeys.

Table 227

Blood counts in cynomolgus monkeys

Table 228

Cynomolgus monkey hematological parameters

Effects on pro-inflammatory molecules

To evaluate the inflammatory effects of ISIS oligonucleotides in cynomolgus monkeys, blood samples were taken for analysis of C-reactive protein and C3 levels on day 84 prior to dosing. About 1.5 mL of blood was collected from each animal and placed in an anticoagulant-free tube for serum separation. The tube was held at room temperature for a minimum of 90 minutes, then centrifuged at 3,000 rpm at room temperature for 10 minutes to obtain serum. C-reactive protein (CRP) and complement C3, serving as markers of inflammation, were measured using a Toshiba 200FR NEO chemical analyzer (manufactured by Toshiba Co., Japan). These results indicate that treatment with ISIS 563580 was highly resistant in monkeys.

Table 229

C-reactive protein levels in cynomolgus monkey plasma (mg/L)

Table 230

C3 levels in cynomolgus monkey plasma (mg/dL)

Determination of Oligonucleotide Concentration

The concentration of full-length oligonucleotides was determined. The method used is a modification of the previously published method (Leeds et al., 1996; Geary et al., 1999), which consists of phenol-chloroform extraction followed by solid phase extraction. An internal standard (ISIS 355868, 27-mer 2′-O-methoxyethyl modified phosphorothioate oligonucleotide, GCGTTTGCTCTTCTTCTTGCGTTTTTT, designated herein as SEQ ID NO: 13) was added prior to extraction. Tissue sample concentrations were calculated using a calibration curve with a lower limit of quantification (LLOQ) of approximately 1.14 μg/g. The results are expressed in μg/g liver or kidney in the table below. The concentration ratio of full-length oligonucleotides in kidney to liver was calculated. After treatment with ISIS 563580, the concentration ratio of full-length oligonucleotides in the kidney to liver was found to be the most optimal compared to other compounds evaluated.

Table 231

Oligonucleotide full length concentration

Example 131: Comparison of antisense inhibition of human ANGPTL3 in huANGPTL3 transgenic mice by ISIS oligonucleotides containing a GalNAc conjugate group and ISIS oligonucleotides without a GalNAc conjugate group

Antisense oligonucleotides comprising GalNAc _{3-7 a} were evaluated for their ability to reduce human ANGPTL3 mRNA transcripts in Tg mice compared to their non-conjugated subjects. A gapmer targeting SEQ ID NO: 1 is described in Table 121 below and Table 121. The symbols for the backbone chemistry columns are as follows: 's' is a thioate ester and 'o' is a phosphate ester.

Table 232

ISIS oligonucleotides

Female and male Tg mice were maintained on a 12 hour light/dark cycle. Animals were acclimatized for at least 7 days in the study facility prior to initiation of the experiment. Antisense oligonucleotides (ASO) were prepared in buffered saline (PBS) and sterilized by filtration through a 0.2 micron filter. Oligonucleotides were dissolved in 0.9% PBS for injection.

Groups of 4 mice were subcutaneously injected with ISIS 563580 at a dose of 5 mg/kg, 10 mg/kg, 15 mg/kg, or 30 mg/kg once a week for 2 weeks. A group of 4 mice was intraperitoneally injected with ISIS 703801 or ISIS 703802 at a dose of 0.3 mg/kg, 1 mg/kg, 3 mg/kg, or 10 mg/kg once a week for 2 weeks. One group of mice received a subcutaneous injection of PBS once per week for 2 weeks. The PBS-injected group served as a control group to compare the corresponding oligonucleotide-treated groups.

RNA analysis

At the end of the treatment period, RNA was extracted from liver tissue for real-time PCR analysis of the measurement of mRNA expression of ANGPTL3 with human primer probe set hANGPTL3_LTS01022. Results are expressed as percent change in mRNA compared to PBS control and normalized to ^{RiboGreen ®.} A value of 0, simply, indicated that the antisense oligonucleotide did not inhibit expression at measurable levels.

The results demonstrate that the conjugated compounds are much more potent in reducing ANGPTL3 expression _{than their non-conjugated compounds, apparently from percent inhibition and ID 50 values.} Conjugated oligonucleotides with mixed backbone chemistry (703802) were more potent at inhibiting expression than conjugated oligonucleotides with full-length phosphorothioate backbone chemistry (703801).

Table 233

% inhibition of ANGPTL3 mRNA in the liver of transgenic mice compared to the PBS control group

protein analysis

Human ANGPTL3 protein levels were quantified using a commercially available ELISA kit (product number #DANL30 by R&D Systems, Minneapolis, Minn.) with transformed plasma samples diluted 1:20,000 using the protocol described by the manufacturer. The results are shown in the table below. A value of 0, simply, indicated that the antisense oligonucleotide did not inhibit expression at measurable levels.

The results demonstrate that the conjugated compounds are more potent in reducing ANGPTL3 expression, apparently from percent inhibition, than their non-conjugated compounds. Conjugated oligonucleotides with mixed backbone chemistry (703802) were more potent at inhibiting expression than conjugated oligonucleotides with full-length phosphorothioate backbone chemistry (703801).

Table 234

% Inhibition of Plasma Protein Levels in Transgenic Mice

Example 132: Resistance of GalNAc Conjugated Antisense Oligonucleotides Targeting Human ANGPTL3 in CD1 Mice

Male CD1 mice (4 animals per treatment group) were subcutaneously injected with ISIS 703802 at various doses for 6 weeks as shown in the table below (1st, 3rd, 5th, 8th, 14th, 21st, 28th, Days 35 and 42) One group of 4 male CD1 mice was injected subcutaneously with PBS for 6 weeks. Mice were euthanized 48 hours after the last dose and organs and plasma were harvested for further analysis.

Plasma Chemistry Markers

To evaluate the efficacy of ISIS 703802, plasma levels of various hepatic and renal function markers were measured at day 44 using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). The results are shown in the table below. ISIS 703802 has been shown to be a tolerable compound even at high doses.

Table 235

Plasma Chemistry Marker Levels in CD1 Mice at Day 44

weight

Body, kidney, liver and spleen weights were measured at the end of the study on Day 44. ISIS 703802 did not significantly change body and organ weights, even when administered at high doses.

Table 236

Weight (g) of CD1 mice after antisense oligonucleotide treatment

Example 133: Resistance of GalNAc Conjugated Antisense Oligonucleotides Targeting Human ANGPTL3 in Sprague-Dawley Rats

The Sprague-Dawley rat is a versatile model used for safety and efficacy evaluation. Male Sprague-Dawley rats were maintained on a 12-hour light/dark cycle and fed ad libitum Purina normal rat chow, Diet 5001. A group of 4 Sprague-Dawley rats was subcutaneously injected with ISIS 703802 at various doses for 6 weeks as shown in the table below (1st, 3rd, 5th, 8th, 14th, 21st, 28th, and no. Days 35 and 42) one group of 4 rats was injected subcutaneously with PBS for 6 weeks. Rats were euthanized 48 hours after the last dose and organs and plasma were harvested for further analysis.

liver and kidney function

To evaluate the efficacy of ISIS 703802 on liver function, plasma levels of transaminase were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY). Plasma levels of ALT (alanine transaminase) and AST (aspartase transaminase) were measured on day 44 and the results are presented in IU/L in the table below.

To evaluate the effect of ISIS 703802 on renal function, plasma levels of albumin, blood urea nitrogen (BUN), creatinine and bilirubin were measured using a homologous clinical chemistry analyzer and the results were measured in g/dL or mg/ It is presented in the table below in dL.

To further evaluate the efficacy of ISIS 703802 on renal function, urine protein and urine creatinine levels were measured, and the ratio of total urine protein to creatinine was evaluated. The results are shown in the table below.

ISIS 703802 has been shown to be a tolerable compound even at high doses.

Table 237

Hepatic and Renal Function Markers in Sprague-Dawley Rat Plasma at Day 44

Table 238

Renal Function Urine Markers (mg/dL) in Sprague-Dawley Rats at Day 44

long-term weight

Body, liver, spleen and kidney weights were measured at the end of the study on day 44 and are presented in the table below. ISIS 703802 did not significantly alter body, kidney, and liver weights even when administered at high doses.

Table 239

Body and Organ Weights of Sprague-Dawley Rats (g)

SEQUENCE LISTING <110> Isis Pharmaceuticals <120> COMPOSITIONS AND METHODS FOR MODULATING ANGIOPOIETIN-LIKE 3 EXPRESSION <130> BIOL0254WO <150> 61/987,467 <151> 2014-05-01 <150> 62/049,230 <151> 2014-09-11 <160> 4914 <170> PatentIn version 3.5 <210> 1 <211> 2126 <212> DNA <213> Homo sapiens <400> 1 ttccagaaga aaacagttcc acgttgcttg aaattgaaaa tcaagataaa aatgttcaca 60 attaagctcc ttctttttat tgttcctcta gttatttcct ccagaattga tcaagacaat 120 tcatcatttg attctctatc tccagagcca aaatcaagat ttgctatgtt agacgatgta 180 aaaattttag ccaatggcct ccttcagttg ggacatggtc ttaaagactt tgtccataag 240 acgaagggcc aaattaatga catatttcaa aaactcaaca tatttgatca gtctttttat 300 gatctatcgc tgcaaaccag tgaaatcaaa gaagaagaaa aggaactgag aagaactaca 360 tataaactac aagtcaaaaa tgaagaggta aagaatatgt cacttgaact caactcaaaa 420 cttgaaagcc tcctagaaga aaaaattcta cttcaacaaa aagtgaaata tttagaagag 480 caactaacta acttaattca aaatcaacct gaaactccag aacacccaga agtaacttca 540 cttaaaactt ttgtagaaaa acaagataat agcatcaaag accttctcca gaccgtggaa 600 gaccaatata aacaattaaa ccaacagcat agtcaaataa aagaaataga aaatcagctc 660 agaaggacta gtattcaaga acccacagaa atttctctat cttccaagcc aagagcacca 720 agaactactc cctttcttca gttgaatgaa ataagaaatg taaaacatga tggcattcct 780 gctgaatgta ccaccattta taacagaggt gaacatacaa gtggcatgta tgccatcaga 840 cccagcaact ctcaagtttt tcatgtctac tgtgatgtta tatcaggtag tccatggaca 900 ttaattcaac atcgaataga tggatcacaa aacttcaatg aaacgtggga gaactacaaa 960 tatggttttg ggaggcttga tggagaattt tggttgggcc tagagaagat atactccata 1020 gtgaagcaat ctaattatgt tttacgaatt gagttggaag actggaaaga caacaaacat 1080 tatattgaat attcttttta cttgggaaat cacgaaacca actatacgct acatctagtt 1140 gcgattactg gcaatgtccc caatgcaatc ccggaaaaca aagatttggt gttttctact 1200 tgggatcaca aagcaaaagg acacttcaac tgtccagagg gttattcagg aggctggtgg 1260 tggcatgatg agtgtggaga aaacaaccta aatggtaaat ataacaaacc aagagcaaaa 1320 tctaagccag agaggagaag aggattatct tggaagtctc aaaatggaag gttatactct 1380 ataaaatcaa ccaaaatgtt gatccatcca acagattcag aaagctttga atgaactgag 1440 gcaaatttaa 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taaagcctgt ctttggtact agtgctcagt tactttcatt aactaaaaaa ggggctactc 180 ttcaaattcc cttctctaaa aagaatgtac tatatcaaaa gggggtaaac actactacgt 240 atacattctg cacttagaaa tccctatatg ttgattttat cattctctta ttcaataaat 300 attgtttcta caatgtgtaa ggcattactg tactaaagca ttataaggaa tataagttaa 360 aaacacatac aaatcttgcc aatcaacagc ttatagtgta ataggggaga gaagctggcc 420 catctatatt ctccctcaac tagcaagtgg atgaaatatc agggtcaata gttataagcc 480 acaaaagctg acagcttaat taagagaagt tttgaaatat gtatttcatg accaataatt 540 acaactgtaa cttttctatt taaagaagga gaaaatttga atttcttctc tagctcaaca 600 tacacttcta taattccatt acatgaacca gagtaaaggg taagatggaa atgaagaata 660 ttttcttacc cttttgtggt tctatattgg acacttaaaa atcatacaca acctaatcaa 720 aagatgtaat tctttaaaaa ggtacgagac caaaattcag aaaatctaga ctataacaaa 780 atttctcaat ttacattatc ttaatatgca attaattttc accagtaaaa tactatagta 840 tgggtacaaa tg cattgatt agttctaatt acaaaaatgg ctaatatata atactgtgta 900 gtgtttatga tacatcagat aatgttctaa gtgctctgaa aatataaact tttaatcttt 960 tatacgaccc tataaaatag gtgttattct cactggagag atgagaaaac aggggttcag 1020 agatgtgaag taatttgacc aaaggtcaca aagctgaaga atatgaaatc cgggattctg 1080 attcaggcag tcttattcca gaatcatgct cttaaccact atggaatact gcctctactg 1140 taactattat acccaaaacc cttaatccta agtcatcaaa aggaagagcc tctattttac 1200 acaatgaaga ggcatttcta agaatagaaa tttagggacg agcacagtgg cttactcctt 1260 taatatcagc actttgagag gctgatatgg gaggttcact tgaagtcagg agttcaaggt 1320 cagcttgggc aacatagtga aacacagtct ctacaaaata tttaaaaatt agctgggtgt 1380 ggtggcatgc atctatagtc tcagctactt gggagacaga gggaggaggc ttgctcgagc 1440 ccaggagttc gtggctatag tgagctatga tcatgccact gcactccagc ctggacaaca 1500 gagcaagacc ctgtctctaa aaaagaaaag aaatttggaa atggtttatt ttgtattaac 1560 aatttataat ttacactgaa atttattatg ataaaacttt tccctgtgtt aaaaagctat 1620 taactttatg aaaaatttct tttaggtaag gttgattata tatacccaca cacatacaca 1680 ggttaaaagt tagtttcatg tgacataata actagcattt tgagcactac ctgtttgccc 1740 agcactgttc taagtgctct acatgtatta ttgttaaatt atcataacac tatgaattat 1800 gtactataat taccccagct ttacagatga ggagactaat ccatggggag gttaagtaac 1860 ttgtccaagg ccagacagct agagccggct tttggaccca caccacagtc tgactccagc 1920 acccatattc ttaacaattt caccatatta atatgtcaag attaagcagt tttaaaggat 1980 gctattttct cacaaatttc ttaatatgaa cactcaataa gaataatcac taatataagc 2040 atttagtatt tttttaacac taagttggaa gcatagtgga acatttattt ttagaaatat 2100 tattaattgg ctgggctcac gcttgtaatc ggctgggctc atgcctgtaa attttgggag 2160 gccaaggtaa aagaattgct tgagcccagt atttccagac cagcatgggc aatacattaa 2220 gacatcatct ttaaaaaaaa aatgttatta atctcctctt tttgttaaat gtatattatc 2280 aaaattgtta ctaagctaac aaacttcaga aaaacttatg atgggcaagc tgcttgtgac 2340 attgaaggta tttaagattc aattctagtt tggtcctaga tgaccacata tccattgttc 2400 cttcaacgag cacatggtaa agagcctaga acacagagac acagaacaca gtggagaaaa 2460 gggagtgaaa tgtctttaat gacacttact atatatggga ttttgtgaca atatacaagg 2520 atggttaaga catataaggt gatgc aaaaa aacatattaa caattatagt gacaaaaaat 2580 gaggagcata taattataca ttgatttata cagagtacca gaggaacaca gcattgagag 2640 ccgtaacacc acctgaggga gtggagaaag gcttcagaga gaaagtgttt tttggaatgg 2700 atcactgttt ccaaaagaac taaagtacag tttgagaaat gcatacttaa ttcattactt 2760 ttttcccctc aactttaata ataaatttac ccaacaaaaa agtttatttt tgacttgtaa 2820 atctcttaaa atcataaaaa agtaaaatta gcttttaaaa acaggtagtc accatagcat 2880 tgaatgtgta gtttataata cagcaaagtt aaatacaatt tcaaattacc tattaagtta 2940 gttgctcatt tctttgattt catttagcat tgatctaact caatgtggaa gaaggttaca 3000 ttcgtgcaag ttaacacggc ttaatgatta actatgttca cctaccaacc ttaccttttc 3060 tgggcaaata ttggtatata tagagttaag aagtctaggt ctgcttccag aagaaaacag 3120 ttccacgttg cttgaaattg aaaatcaaga taaaaatgtt cacaattaag ctccttcttt 3180 ttattgttcc tctagttatt tcctccagaa ttgatcaaga caattcatca tttgattctc 3240 tatctccaga gccaaaatca agatttgcta tgttagacga tgtaaaaatt ttagccaatg 3300 gcctccttca gttgggacat ggtcttaaag actttgtcca taagacgaag ggccaaatta 3360 atgacatatt tcaaaaactc aacatatttg atcagtcttt ttatgatcta tcgctgcaaa 3420 ccagtgaaat caaagaagaa gaaaaggaac tgagaagaac tacatataaa ctacaagtca 3480 aaaatgaaga ggtaaagaat atgtcacttg aactcaactc aaaacttgaa agcctcctag 3540 aagaaaaaat tctacttcaa caaaaagtga aatatttaga agagcaacta actaacttaa 3600 ttcaaaatca acctgaaact ccagaacacc cagaagtaac ttcacttaaa gtaagtagaa 3660 aataaagagg gttcatgttt atgttttcaa tgtggatctt ttaaaaaaaa tatttctaag 3720 gcatgccatt tgaaatactt tgttgcattg ttgaaatact tttttttcca agaaaaataa 3780 tctccagaaa ataaaatttc ctattataat ttcaagttag ttttttgttt ccctaatgtt 3840 atatatgaaa acactgaaaa tttgcatttt atatgaaaat tacaaatcgg ttaaattata 3900 caatctagaa cactatgtca ttacactatt gtaaattact gaaggtaagt aaaaagttaa 3960 aaaaaattta aaactattct ccagtgttta aaacagatta aataatacag taaatggaaa 4020 agatttattc atatgaaaat atgctgggct ttttctttta attgaagttc agaaaatcaa 4080 attttagaga tagtacaatt taaataaaat gttaaggaca aaaatatgtg ctatttgaaa 4140 gaagcataca aggggaagga attgccaata ttcatttttc aaatccatta ttagtttaaa 4200 aatttagatt atgatagtgt tacaggaaat taatag aaaa gaaagaggaa agcaacttat 4260 aaccaaccta ctctctatat ccagactttt gtagaaaaac aagataatag catcaaagac 4320 cttctccaga ccgtggaaga ccaatataaa caattaaacc aacagcatag tcaaataaaa 4380 gaaatagaaa atcaggtaag tcagtatttt aatggtatgt cccatctttc acacaggtct 4440 gtaaaaacac tgaatcctaa aattatttac aagctttaac tggatcatga gtaaaattat 4500 cacatcagca taactgttaa aattgcaggc tctgaagcta ataaactacc tgcatttaaa 4560 ccatggctct aaaactttgt gtgaccttga ataaattact tcaccccttt atctctcagt 4620 ttcctcacat atactacaaa gataataaca gaacttatag gattattgta agaaaaaaaa 4680 ttaattcata gcagccaatg tcatcttact aaaattcaaa ttagatcatg tttctctttg 4740 ctcaaaacca cacaatagct ttccatttca ctcatattgg ctctttagac caagattacc 4800 caacccttcg tcatctcact gacttcacct cctctactct agttattctg accgctttac 4860 cagtattcaa acacatcaaa catactgcca cctcaaagcc tttgcccttg ttgtttcctc 4920 taactggaac gctcttctgc cctggtatct acgtggccca ctctctgatt tcccttaggg 4980 tcgttatcaa acaaaaaatt cccaatgaag acttacaagg tcacttaacc aaaaatcaca 5040 accgcctggt cccatccctg aaaacttcta cttccttagc t acttttctc ctgcacactc 5100 acctttattt aacataacat aaattttagt tatttatctc ttctattcct gcactaaaat 5160 gtaagctctg tgaatacagg gattttttcc attatcttca tattttccat tatttgtata 5220 tactccagaa tatagaatac tgtatggcac acagtaggca tttctgttga attaataaat 5280 gtaatgtcat attcacacag aagcgtgtgc tatgattatt attacttgga ttactagaaa 5340 tagtgtgcct cataattaaa ggtcaacatt caacaatgta attaatctac aatgtaaaca 5400 tctggtgaag tgacagaggg aagcacttgt ttagaaaaaa gctatgtcag aatccatgta 5460 ttctaatatg cagtacaata gtttaaaaat attaataata ctctcaaaca gctattcaag 5520 aggattcaaa aaacataata taaactcaga gaaactggta aacaaaatca ttttcaagag 5580 atataaaaca aatattatta ccaatttcca ctaaacaaac ataatgttag tagtgctgct 5640 aaaaggtttt ttatcaacta cttttggttt ccatactttc cttcttatga tgttattatt 5700 ctaaattctt ttcaattata tcttttacta tgattaaatg aacctgctcc ccaaagcaaa 5760 atgttactat agtaatatac attgtgtcta aaaataaaaa tgtgtgaaga aaccaaaaca 5820 atgaatttct gagttggaag aagagttaga tcatttaact ttctcatatt taaattaaaa 5880 aaacaaaact ctaaaaattt aagtaacttt aagatcacat agttact tag tagaaaagag 5940 taatacccag caagcaaact ttacaataga tccttttaaa taaggtccta ggaaatatca 6000 ttcatgccag catcaaaaaa ctaacactaa taatgcaaga tattatatat tctgcttttc 6060 ttactgtcaa tgagaaaaac tatcattcaa taaattgcaa acccaacaca cttaaataaa 6120 aataaaatgt tactgctaaa ctaacgataa actactgaat atatagaaag taagcaaaca 6180 aacttgccaa cctgccaaca tctacagata tgtttacagg tcaaaaatta tcaaattatc 6240 aagaaagcct ggttcaaatt atgtattatg tctttatcac aggtctgaag atcagtaaga 6300 cctaaaactg aaaattatta aacttaaaat ctgaacagaa tatcaaatat attttattca 6360 tataaataaa agaatacatt acaatattct aagcaaagca gtctctactt ttggccttgc 6420 tctgttttcc gaccaatgtc tgcttttttg ccttgcttta tttttttatc ttattaaata 6480 atgtccctga ttaaatattt tgagaacagg taatctgtac aatctgaata acactgttta 6540 tctaaatatc aaacaccgtt ataacattat gaactgaaag acaaactgta cttctgacat 6600 ccttactcag atttccccta attgtatatt cagtatcatt ttaaaaaaca gatttatatt 6660 cttttatcag ctcagaagga ctagtattca agaacccaca gaaatttctc tatcttccaa 6720 gccaagagca ccaagaacta ctccctttct tcagttgaat gaaataagaa at gtaaaaca 6780 tgatggtaag acactttggt gggtttcctt cttgaagcta ttattatcaa attccctatt 6840 cttaggactt gttctagact aaaagatagt taagagatat ccatcaaata caatgtatca 6900 acctaaactg gatgctgggg ttctttttac accctataaa agacatacct aagacaatca 6960 gagaaataca aatatggact tgattattag ataatataga aggtttatta attttcttag 7020 atgtgatcat ggtattgcag ttttaaagga gaacaatctc ctgtttaaga gatacatgct 7080 gaaatattta cggagttaaa ggtcactgga ctccagactg gtgatagaac aagactctgt 7140 ctctaaaaaa taattaattt tttaaaagaa aatagtttgg taagatgatt cttacattct 7200 taaataacac gccatctaag aaaaatgctt taacataaac attactgaaa aaatgctaca 7260 tttgccacaa cttcataaaa tgtcaagtga aatctcaagc tccaaagata ttattcctat 7320 tactaaatct gatgtaataa cattttattg attctaggca ttcctgctga atgtaccacc 7380 atttataaca gaggtgaaca tacaagtggc atgtatgcca tcagacccag caactctcaa 7440 gtttttcatg tctactgtga tgttatatca ggtaaaacct gtctaaggag aatagacagt 7500 agttagttca acttactcat tacgtattag gaagattaac ctggttatca ttgttttata 7560 catatatata tgaaatatat atgagtattc gtataaatat aatactttta ccttgttt at 7620 gtatttactc aatattctcc ttttcctcta aaataatctg aagtgactat tatcaataag 7680 tttactatgc caaaattcat taattgcctt tcacttaact tttgggacca taataaataa 7740 taaaatgtat tgccataaca ttaataaact accttacaaa accaccaatt aaaatcaaac 7800 aaacaaaaaa gtgttattta catctgtcaa cataaatcta ctaaaaatac atgatttcat 7860 tcattatatt caggtagtcc atggacatta attcaacatc gaatagatgg atcacaaaac 7920 ttcaatgaaa cgtgggagaa ctacaaatat ggttttggga ggcttgatgg taaggggact 7980 acattcaatc attcattcac ttgctaatct acaaatattt actgagaacc tcttatggac 8040 caggtattag gaaaagtagt aacgaacgag aagcagtctc agccttcata taatttatta 8100 tcaaacaatt acacatttgt tagtaaatta cacttattac aactgttatt atttgaatta 8160 tatttatcac aattacatgt ctgttcttaa atatacttat cacaatttaa ttccacggct 8220 tacaatgatc ataactataa ttattaaaga caattttgat taaatgttat gtcataagta 8280 gtaactgtta caaataagct gtgaaaagaa ccactcctag cattagtcac tctattctct 8340 cattaacgtt ttacatatca attaattgga agttaaaagg accaggaaac tcagacatac 8400 agtatacatt ttaaaatttc aattatttaa atataatata tagaatgtat ggcttataat 846 0 gaattagtta actcaatgca aattattcta ttttgattac aaatagtaaa ataagcaaga 8520 taaaataaca gatgtttaaa atccaaaaag cacatacaaa aatccatgaa tgatgtctaa 8580 gtactcactt ataaagtaga agacattcat tattatatca aatttttaaa tgctcagtac 8640 tatttgacca tttaaaaatt ttgtattcaa actaccagtg aaagccctac ctagaaggta 8700 tactcagtga taagttttgt agctccaaat cttctaatag tgagtgtaac cccaaaataa 8760 aaggctgaca ggtaagtcga gaatactcac ttaattctgg taagaaagca acccatttgt 8820 acttgtattt accagcaatc cttaaaatga agcttcctac taactcaata gcaataagac 8880 aatagtgaat gtttaatgaa aacagtattt tataaatact ttaataaaaa ggattgtgat 8940 gaagaacaat ctatttatat ttgttatttg tttttaattc caataaaaat aatttttaaa 9000 attacagaaa aaagttatta agaaccatgc ttttaaattt aaaatgattt tttaaattta 9060 ttcctgtctt tttctacaaa gaaagcatac attaagcaaa taccaaaggc caggtttaca 9120 tttgaagaaa gtgacattat tattactcaa gtctctagga atacttaaca catctcttga 9180 ctgtatatgg atgttaataa atagctgaca gtaaagttta tccatataaa gacttgcaaa 9240 tattcctcta ccaatgacga gactttaaaa tatctataat aatgtaacac atttcactgg 9300 tgaa acatgt cttgtcatat gcattataga aaggataatc agactttcag ttatattaat 9360 atttttaaca tttttgtgca catagctatc ttcaataaaa ttgttttaaa aggtattatt 9420 ttaagataca ctaaaatgat caagggattc aagactaaac aactcaatta gttgcaccaa 9480 taaaaaacac ttaaaaaaac tgtcagtgtc caacctgtac ttaataactc acagattttt 9540 aaaacttttc ttttcaggag aattttggtt gggcctagag aagatatact ccatagtgaa 9600 gcaatctaat tatgttttac gaattgagtt ggaagactgg aaagacaaca aacattatat 9660 tgaatattct ttttacttgg gaaatcacga aaccaactat acgctacatc tagttgcgat 9720 tactggcaat gtccccaatg caatcccgga aaacaaagat ttggtgtttt ctacttggga 9780 tcacaaagca aaaggacact tcaactgtcc agagggttat tcaggtatct ttttctgata 9840 ccaatacttt attttcatat cttcaaagta tcttcccaca ttattagcta ttatctgcaa 9900 tgacaacttt taaaaatccg aatcccaaat aagcgttttc tctctagacg aaaacctctt 9960 aactataatg aaagtgttca ttctagttca atcaggtatt ttacctctaa tcttcctcag 10020 attttctatt ttttggtagt gtatagatta tttatacaga ttatttaaaa ttgggactta 10080 tacagattat ttaaaactgg gatacatgca tctaaaacac tgtaatattt ataagaaagg 10140 aagataa act tacggggaaa tacagtaaca gtaactacat acgagtctgt acccattaaa 10200 ttgcatatct atctccttta ggaggctggt ggtggcatga tgagtgtgga gaaaacaacc 10260 taaatggtaa atataacaaa ccaagagcaa aatctaagcc agagaggaga agaggattat 10320 cttggaagtc tcaaaatgga aggttatact ctataaaatc aaccaaaatg ttgatccatc 10380 caacagattc agaaagcttt gaatgaactg aggcaaattt aaaaggcaat aatttaaaca 10440 ttaacctcat tccaagttaa tgtggtctaa taatctggta ttaaatcctt aagagaaagc 10500 ttgagaaata gatttttttt atcttaaagt cactgtctat ttaagattaa acatacaatc 10560 acataacctt aaagaatacc gtttacattt ctcaatcaaa attcttataa tactatttgt 10620 tttaaatttt gtgatgtggg aatcaatttt agatggtcac aatctagatt ataatcaata 10680 ggtgaactta ttaaataact tttctaaata aaaaatttag agacttttat tttaaaaggc 10740 atcatatgag ctaatatcac aactttccca gtttaaaaaa ctagtactct tgttaaaact 10800 ctaaacttga ctaaatacag aggactggta attgtacagt tcttaaatgt tgtagtatta 10860 atttcaaaac taaaaatcgt cagcacagag tatgtgtaaa aatctgtaat acaaattttt 10920 aaactgatgc ttcattttgc tacaaaataa tttggagtaa atgtttgata tgatttattt 10980 atgaaaccta atgaagcaga attaaatact gtattaaaat aagttcgctg tctttaaaca 11040 aatggagatg actactaagt cacattgact ttaacatgag gtatcactat accttatttg 11100 ttaaaatata tactgtatac attttatata ttttaacact taatactatg aaaacaaata 11160 attgtaaagg aatcttgtca gattacagta agaatgaaca tatttgtggc atcgagttaa 11220 agtttatatt tcccctaaat atgctgtgat tctaatacat tcgtgtaggt tttcaagtag 11280 aaataaacct cgtaacaagt tactgaacgt ttaaacagcc tgacaagcat gtatatatgt 11340 ttaaaattca ataaacaaag acccagtccc taaattatag aaatttaaat tattcttgca 11400 tgtttatcga catcacaaca gatccctaaa tccctaaatc cctaaagatt agatacaaat 11460 tttttaccac agtatcactt gtcagaattt atttttaaat atgatttttt aaaactgcca 11520 gtaagaaatt ttaaattaaa cccatttgtt aaaggatata gtgcccaagt tatatggtga 11580 cctacctttg tcaatactta gcattatgta tttcaaatta tccaatatac atgtcatata 11640 tatttttata tgtcacatat ataaaagata tgtatgatct atgtgaatcc taagtaaata 11700 ttttgttcca gaaaagtaca aaataataaa ggtaaaaata atctataatt ttcaggacca 11760 cagactaagc tgtcgaaatt aacgctgatt tttttagggc cagaatacca aaatggct cc 11820 tctcttcccc caaaattgga caatttcaaa tgcaaaataa ttcattattt aatatatgag 11880 ttgcttcctc tatttggttt ccttaaaaaa aaaaaaaact ctcataggac atgtttcatt 11940 ttgttccttt caggagtagt aaattagacg ttttccccat ataaagcttt tttctaccag 12000 aaagatactt ctggtagaag aagagaaagg agctctttat ggttcacacg actgtctcct 12060 gtcctaacta ctttgcttaa agtgctcaaa ttccatcact actcacagtt gtctaatcta 12120 agtctaatcc cctttgatct ctcagactac cttccctttt atctctctac tacttaataa 12180 taagaatatc tttttttcaa acttgacctt cattttgctt tcacaatact atactctcca 12240 tggattatcc cttatctgaa tccatcttta taaccctatt cctttctcat atttagtact 12300 gtgggccaat ggacaacctt caatcatctt ttctacactg accctcagac attctatctg 12360 ctctcacgga ctcctttatt taccatgaat aaagttccaa aatctacata ttcatcccaa 12420 gtctctttcc agttcccctt cttacattgc ctatttgcca tttctccctt caatacccta 12480 tacttcactc aaattcaaca taccaaaaat aaaaggccag gcacggtggc tcacacctgt 12540 aatcccagga ctttgggagg ctgaggcagg tggatcacct gaggtcagga gtctgaccag 12600 cctgaccaat atggtgaaac cccgtctcta cctaaaatac aaaaattagc caggcgtggt 12660 ggcatgtgcc tacagtccca gctactcaag aggctgagac aggagaatcg cttgaaccca 12720 ggaggcggag gttgcagtga gctgagatca caccaatgca ctgggtgaca gaacaagact 12780 gactcaaaaa aaaataaata acaaattccc cagcccctta ctgctactgc tatccctttc 12840 tacccacctt tccctccttt atactctttc acaccatctt cctcacttct ttatatccat 12900 taatatgacc agcatgttcc cagtcacaga agcctggaac ccggaagaca tctctggctt 12960 ttcactcaac tttgtaaact acctcttttg tatcataagc caccaagttc aatacaatct 13020 tctcttgaaa cgtctcttaa tcttataagc tttcttcccc aaagactgtc tttaacttca 13080 gtgctagatt atataagtag tctcttcttt cctcctaatt ttacgtccct ctccctgtca 13140 atcttccgaa aaggaatttg taatggctgc atactgcctg gacctgtgtc tttcaagctc 13200 atctactgaa gaagtgtaac agcagaggat aatgaaaaca aatcctttta ttgtctggag 13260 ttacaggata aaataactta ctatatgcac aaatttcttt cagaatctta ttgggatctt 13320 agcttaaatc ttagaatttt actttaaaat tgcatatgaa ttgtgtttta tactttataa 13380 atctatcttc atgcctgcag tggctcacac ctgtaatcca agtattttgg gaggccgggg 13440 cgggagtact gcttgaaccc aggagtccaa gatcagcctg tgc aacacgg caaaactcca 13500 cctctacaaa aaatacaaaa attagctggg tgtggtggca ggtgcctgta atcccaacta 13560 cttgggaggc tgaggcagga gaatcgctgg agctactgggaa gtcaagcctgcataact cagcgagtactg actaacta 13620 cagcgagtactg ttcatataaa aatatttatt tccccccaag tatttaaaaa gtgatgttcc tagaagatgt 13740 gtcatgttta tcctgtgaac ttgcacgtcc catatagttg tgccctcaca agaatgtata 13800 ctccaattag aagagtatag gcctttggaa caaaatccaa aatccttcca ggttttcaaa 13860 agtctctatc acctggcccc aaattacctc ttcagccatt ttttcccaca acacaaacct 13920 tctgctctca tcaaaacgat ctacttttaa acatctcctc tatgccttcc taatgtattc 13980catttgtcta gaatgtctcc 14000 <210> 3 <211> 13000 <212> DNA <213> Macaca mulatta <220> <221> misc_feature <222> (4504)..(4523) <223> n is a, c, g, or t <220> <221> misc_feature <222> (5370)..(5795) <223> n is a, c, g, or t <220> <221> misc_feature <222> (7498)..(7517) <223> n is a, c, g, or t <220> <221> misc_feature <222> (9552)..(9895) <223> n is a, c, g, or t <400> 3 cccagctact gaggaggctg aggcaggaga atggcgtaaa cccgggaggc ggagcttgca 60 gtgagctgag atctggccac tgcaccccag cctgggcaac agagcaagac tccgtctcaa 120 aaaacaaaac aaaacaaaac aaaacaaaac aaaacaaaga atgtagtata tcaaaaagag 180 gtaaccagta ctacatatac attctgcact tggaaaatcc ctatatgttg attttatcat 240 tctcttagtc attcaataaa cattgcttct acaatgtgtg aggcattact gtactaaagc 300 attataagga atatacatta aaaacacata caaatcttgc caataaacgg cttatagtat 360 aataggggag agaagttggc ccatctatct tctccttcaa ctagcaagta gatgaaatat 420 cagggtcaat agttgtaagc cataaaagct gacagcttag ttaagagaag ttttgaaata 480 tgtatttcat ggccaaataa ttacaactgt aacttttcta tttaaagaag gagaaaattt 540 gaatttcttc tctagctcaa catacacttt tataattcta taacatgaac cagagtaaag 600 ggtaagatgg aaatgaagaa tattttctta cccttttgtg gttccatatt ggacacttaa 660 aaatcataca caacctaatc aaaagacata attcattaaa aaggtacgag accaaaacta 720 aaaaaatcta gactataaca aaatttccca atttacatta tcttaatatg caattaattt 780 tcaccagtaa aatactatac tatgggtaca aatgcattga ttagttctaa ttacaaaaat 840 ggctaatata ta atattatg tagtgtttat gacacaccag gtaatgttct aagtgctttg 900 aaaatataaa cttttaatct ttatacaacc ctataaaata ggtgtcattc tcactggaca 960 gatgagaaaa caggggttca gaaatgtgaa gtaatttgac caaaggtcac aaaactgaag 1020 aatatcaaat ccaggattct gattcaggca gtcttgttcc agaatcatgc tcttaaccac 1080 tatggaatac tacctgtatt atagtattac atatactgta ctataactat tacacccaaa 1140 acccttaata ctacctgtac tataactatt atacccaaaa tcctgagtca tcaaaaagaa 1200 gagcctctat tttatacaat gaagaggcat ttctaagaat agaaatttag ggacgagcac 1260 agtggcttac tcctgtaata tcagcacttt gagaggctga tgtgggaagt tcacttgaag 1320 tcaggagttc gaggccagct tggacaacat agtgagaccc actctctaca aaatatttaa 1380 aaattagctg ggcgtgttgg catgcatcta tagtctcagc tactggggag gctgagggag 1440 gaggctcact caagcccagg aattcgtggc tatagtgagc tatgatcatg ccactgtact 1500 ccagcctgga caacagagca agaccctgtc tctaaaaaaa agaaaagaaa agaaatttgg 1560 aaatggttca ttttgtagta acaatttata atttacactg aaattcatta tgataaaact 1620 tttccctgtg ttaaaaagat attaacttta tgaaaaattt attttaggta aggttgatta 1680 tatatactca cacacataca caggttaaaa gttagtttca tgtgacataa taactagcat 1740 tttgagcact acctgtttgc ccaacactgt tctaagtgct ctacatgtat tattgttaaa 1800 ttatcataac actatgaatt atgtactata attaccccag ctttacagat gaggagacta 1860 atccaagggg aagttaagta acttgtccaa ggtcagacag cgagagctgg ctttggaccc 1920 cactacagtc tgactccagc acccatattc ttaacaattt caccatatta atatgtcaag 1980 attaagtagt tttaaaggat actattttct cacaaagttc ttaatatgaa cacttaataa 2040 gaataatcac taatattagt atttttcttt ttaacactaa gttggaagca cagtggaaca 2100 tttattttta gaaatattat taattggctg ggctcacgcc tgtaattgac tgggctcatg 2160 tccgtaaatt ttgggaggcc aaggcaaaag aatcgcttga gcccaatagt tccagaccag 2220 cataggcaat acattaagac atcgtcttta aaaaaacaaa gttattaatc tcttcttttt 2280 gttaaatgta tattatcaaa attgttacac taagctcaca aacttcagaa aaacttatga 2340 tgggtaagtt gcttgtgaca ttgaaagtat ttaagattca attctgtttg atcctagaag 2400 accacatatc cattgttcct tcagtaaaca catgataaag agcctagaac agagagacag 2460 agaacacagt ggagaaaggg gagtgaaatg tctttaatga cacttactat atgtgggatt 2520 ttgtgataat atacaaggat gatta agaca tataaggtga tgcaaaaaaa ccatattaac 2580 aattatagtg acaaaaataa ggagtacata attatacgtt gatttataca gagtaccaaa 2640 ggaatacagc attgagagct gtaccactac ccgagggaat ggagaaaggc ttcagagaga 2700 aagtgttttt tgggatggat caatgtttcc aaaagaaccg aagtgcagtt tgagaaacgc 2760 atacttaatt aattcttttc ctcaacttta ataataaatt tactcaacaa aaaagtttat 2820 ttttgacttg taaatctctt aaaatcataa aaaagtaaaa tcagctttta aaaacaggta 2880 gtcaccatat cattgaatgt gcagtttata atacagcaaa gttaaataca atttcaaatt 2940 acctattaag ttagttgctc atttctttga tttcatttag cattgatgta actcaatgtg 3000 gaagaaggtt acattcgtgc aagttaacat ggcttaatga ttaactatat tcacctgcca 3060 accttgcctt ttctgtggca aatattggta tatatagagt taagaagtct aggtctgctt 3120 ccagaagaac acagttccac gttgcttgaa attgaaaatc aggataaaaa tgttcacaat 3180 taagctcctt ctttttattg ttcctctagt tatttcctcc agaattgacc aagacaattc 3240 atcatttgat tctgtatctc cagagccaaa atcaagattt gctatgttag acgatgtaaa 3300 aattttagcc aatggcctcc ttcagttggg acatggtctt aaagactttg tccataagac 3360 taagggccaa attaatgaca tatttcaaaa actcaacata tttgatcagt ctttttatga 3420 tctatcactg caaaccagtg aaatcaaaga agaagaaaag gaactgagaa gaactacata 3480 taaactacaa gtcaaaaatg aagaggtaaa gaatatgtca cttgaactca actcaaaact 3540 tgaaagcctc ctagaagaaa aaattctact tcaacaaaaa gtgaaatatt tagaagagca 3600 actaactaac ttaattcaaa atcaacctga aactccagaa catccagaag taacttcact 3660 taaagtaagt agaaaataaa gagggttcat gtttatgttt tcaatgtgga acattaaaaa 3720 aaaatatttc taaggcatgc tacttgaaat actttgttgc attgttggaa tacttttttt 3780 tccaagaaaa ataatctcca gaaaataaaa tttcccatta taattttgtt agtttttgtt 3840 tccctaatgt tatatatgaa aacactgaaa ctttacattt tatatgaaaa ttacaaattg 3900 gttaagtaac acaatctaga acactatgtc attacactat tgcaaattac tgaaggtaag 3960 taaaattttt taaaaattta aaactattct ctagtgttta aaacagatta aagtaataca 4020 gtaaatggaa aagatttatt cctattaaaa tatgctgggc tttttctctt aattgaagtt 4080 cagaaaaaca aatcttagag atagcacaat ttaaataaaa tgttaagaac aaaaatacat 4140 gctatttgaa agaagcatac aaggggaagg aattgccaat attcattttt caaatccatt 4200 attagtttaa aaatttagat tatgatagtg ttacag gaaa ttaaaactct aatagaaaag 4260 aagaaagtaa cttataacca atctaatctc tatattcaga gttttgtaga aaaacaagat 4320 aatagcatca aagaccttct ccagactgtg gaagaacaat ataagcaatt aaaccaacag 4380 cacagtcaaa taaaagaaat agaaaatcag gtaagtcaat attttaatgg tatgtcccat 4440 ctttcacaca ggtttgtaaa acattgaatc ctaaaattat ttacagcttt aactggatca 4500 tgannnnnnn nnnnnnnnnn nnngaaaata attaattcat agcagccaat gtcatcctat 4560 taaaattcaa attagatcat gtttctcctt gctcaaaacc acacaatagc tttccatctc 4620 actcatattg gctctttaga ccaagattac ccgacccttc atcatctcac tgacttcacc 4680 tcctctactc tagtcatttt gatcgcttta ccggtattca aacacatcaa acatgctgcc 4740 acctcgaagc ctttgccctt gttgtttcct ctatctggaa tgctcttttg tcctggtatc 4800 tacagggccc actctttgat ctcccttagg gtcgttatca aacataaaat tctcaatgaa 4860 gatttccaag gtcacttaac caaaaattac aactgcctgg tcccatccct gaaaacttct 4920 acttccttaa ctacttttct cctgcacact cacctttatt taacataaat tttagttatt 4980 tatctcttct attcctgcac taaaatgtaa gctctgtgaa taaaggggtt tttttccatt 5040 atcttcatat tttccattat ttgtatatac tccagaatat a gaatactgt atggcacata 5100 ctaggcattt ctgttgaatt aataaatgta atgtcatatt cacacagagt gtgtgctata 5160 attattatta cttggattac tagaaatagt gtgcctcata attaaaggtc aacattcaac 5220 aacgtaatta atctacaatg taaacaactg gtgaagtgag gggggaagca cttgtttaga 5280 ataaagctat gtcagaatct aagtattcta atatacagta aatagttaaa aaatattaat 5340 aatactctta aacagtcatt caagaggatn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 5400 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 5460 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 5520 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 5580 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 5640 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 5700 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 5760 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnntaacg ttaaaatcac atagttactt 5820 agtagaaaag agtaatatcc agcaagtaaa ctttacaata gatcctttta aataaggtcc 5880 taggaaatgt catttatgcc aggatcaaaa aactaacact cataatg caa gatattatat 5940 attctgcttt tcttactgtc aatgagaaaa actatcattc aataaattgc aaacccaaca 6000 cacttaaata aaatgttatt gcaaaactaa cgataaacta ctgaatatat ggaaagtaag 6060 caaataaact tgccaacctg ccaacatcta cagatgtgtt tacaggtcaa aaattatcaa 6120 attatcaaga aagccaggtt caaattatgt attatgtctt taaagcaggt ctgaagatca 6180 gtaagaccta aaactgaaaa ttattgaact taaaatctga acagaatatc aaacatattc 6240 tattcatata aaaaaaaata cattacaaca ttctaagcaa agcagtctct acttttggcc 6300 ttgttctgtt ttctgaccaa tgtctgcttt tttgccttgt tttatttttt taatttatta 6360 aataatgtcc ctgattaaat attttgagaa caggtaatct gtacaatctg aataacactg 6420 tttatctaaa tatcaaacac cataacatta tgaactgaaa gacaaactgt acttttgaca 6480 tccttactca gatctcccct aattatgtat tcagtatcat tttaaaaaac agatttatat 6540 tcttttatca gctcagaatg actaatattc aagaacccac agaaatttct ctatcttcca 6600 agccaagagc accaagaact actccctttc ttcagctgaa tgaaataaga aatgtaaaac 6660 atgatggtaa gacactttgg tgggtttcct tcttgaagct attatcaaat tccctattct 6720 taggacttgt tctagactaa aagattaaga gatagccatc aaatacaatg ga tcatccta 6780 aattggatcc tggggttctt cttctaccct ataaaagata tacctaagac aatcacagaa 6840 atataaatat ggacttgatt attagataat atagaaggtt tattaatttt cttagacgtg 6900 atcatggtat tgcaatttta aaggagaaca acctcctgtt taagagatac atgctgaaat 6960 atttaaggag ttaaaggtca ctggactcct ggctgggtga cagaacaaga ctctgtctct 7020 aaaaaaataa ttttttaaaa gaaaatagtt tggtaagaat gattcttaca ttcttaaata 7080 acatgccatc gaagaaaaat gctttaacat aaacattact gaaaaaaatg ctacatttgc 7140 cacaatttca taaaatgtca agtgaaatct caagctccaa agacattatt cctattacta 7200 aatctgatgt aataacattt tattgattct aggcattcct gctgattgta ccaccattta 7260 caatagaggt gaacatataa gtggcatgta tgccatcaga cccagcaact ctcaagtttt 7320 tcatgtctac tgtgatgttg tatcaggtaa aacctgtcta aggagaatag acagtagtta 7380 gttcaagtta ctcagtatgt attaggaaga ttaacctggt tataattgtt ttatacatat 7440 atatatatga aatatatgta acatgagtat tagtataaat ctaatacttt tatcttgnnn 7500 nnnnnnnnnn nnnnnnnatg gatcacaaaa cttcaatgaa acgtgggaga actacaaata 7560 tggtttcggg aggcttgatg gtaaggtgac tacattcaat cattcattca tttgctaa tc 7620 tacaaatatt tactgagaac ctattgtgga ctgggcatta ggaaaagtag taatgaagaa 7680 gcagcagtct cagccttcat ataatttatt atcaaacaat tacacatttg ttagtaaatt 7740 attacaactg ttataatttg atttatattt atcacagtta catgtctgtt cttaaatata 7800 cttatcacaa tttaatttga tggcttacaa tgatcataac tataattatt aaagacaatt 7860 ctgattaaat gttatataag tagtaactgt tacaaataag ctgtgaaaag aaccactcct 7920 agcattagtc actctattct ctcattaacg ttttacgtat caattaattg gaagttaaaa 7980 ggaccaagaa actcagacat acagtataaa ttttaaaatt tcaattattt aaatgtaata 8040 tatagaatgt gtggcttata atgaattagt taactcaatg caaattattc tgattacaaa 8100 ataatgttaa ataagcaaga taaaataaca gatgtttaaa atccaaaaag cacatacaaa 8160 aatccatgaa tgatgtctaa gtactcactt ataaagtaga agacattcat tattatatca 8220 aatttttaaa tgctcagtac tatttgccca tttaaaaatt ttgtattcaa attacccgtt 8280 aaagccctac ctagatggta tactccgtga tatgttttgt agctccaaat cttctaatag 8340 tgagtgtaac cccaaaataa aaggctgaca ggtaagttaa gaatactcac ttaattctgg 8400 taagaaacta acccatttgt acttgtattt accagcaatc cttaaaatga agcttcctac 846 0 taactcaata gcaataagac aacagtgact gtttaatgaa aacagtaatt tattaagcat 8520 tttactaaat actttaataa aaaggattgt gataaaaagt ctatttatat ttgttattta 8580 tgtttttaat tccaaataaa aataattttt aaaattatag aaaaaagtta ttaagaacca 8640 tgctttttca tttaaaatga tttttaaaat ttattcctgt ctttttctac aaagaaaaca 8700 tatattaagc aaataccaaa ggccaggttt agacttatta ttactcaagt ctctgggaat 8760 acttaacaaa aatctcttga ctgtatatgg atgttaataa atacctgata gtaaagttta 8820 tccatataaa gacttacaag aaaatattcc tctaccaatg acgagacttt aaagtatcta 8880 taataatgta acatatttca ctggttaaat atgtcttgta atatgcatta tagaaaggat 8940 aatctgactt tcagttctat taatattttt aacattattg tgcatataga tatcttcaat 9000 aaaattgttt taaaaggtat tattttaaga tactctaaaa tgatcaaggg attcaagact 9060 aaacaactca attagttgca ccaataaaaa acacttaaaa aaacagtgtc caacctgtag 9120 ttaataattc acagattttt aaaacttttc ttttcaggag aattctggtt gggcctagag 9180 aagatatact ccatagtgaa gcaatctaat tacgttttac gaattgagtt ggaagactgg 9240 aaagacaaca aacattatat tgaatattct ttttacttgg gaaatcacga aaccaactat 9300 acgc tacatg tagttaagat tactggcaat gtccccaatg caatcccgga aaacaaagat 9360 ttggtgtttt ctacttggga tcacaaagca aaaggacact tcagctgtcc agagagttat 9420 tcaggtatct ttttctgata acaatacttt attttcatat cttcaaagta tctgcccaca 9480 ttattagcta ttatctgcaa tgacaacttt taaaaatccg aatcccaaat aagcgttttc 9540 tttctagatg annnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 9600 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 9660 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 9720 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 9780 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 9840 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnacact 9900 ataatattta ttaaaaagga agataaactc atggggaaat acagtaacta catacgagtc 9960 tgtacccact aaactgcgta tctatctcct ttaggaggct ggtggtggca tgatgagtgt 10020 ggagaaaaca acctaaatgg taaatataac aaaccaagaa caaaatctaa gccagagcgg 10080 agaagaggat tatcctggaa gtctcaaaat ggaaggttat actctataaa atcaaccaaa 10140 atgttga tcc atccaacaga ttcagaaagc tttgaatgaa ctgaggcaaa tttaaaaggc 10200 aataaattaa acattaaact cattccaagt taatgtggtt taataatctg gtattaaatc 10260 cttaagagaa ggcttgagaa atagattttt ttatcttaaa gtcactgtca atttaagatt 10320 aaacatacaa tcacataacc ttaaagaata ccatttacat ttctcaatca aaattcctac 10380 aacactattt gttttatatt ttgtgatgtg ggaatcaatt ttagatggtc gcaatctaaa 10440 ttataatcaa caggtgaact tactaaataa cttttctaaa taaaaaactt agagacttta 10500 attttaaaag tcatcatatg agctaatatc acaattttcc cagtttaaaa aactagtttt 10560 cttgttaaaa ctctaaactt gactaaataa agaggactga taattataca gttcttaaat 10620 ttgttgtaat attaatttca aaactaaaaa ttgtcagcac agagtatgtg taaaaatctg 10680 taatataaat ttttaaactg atgcctcatt ttgctacaaa ataatctgga gtaaattttt 10740 gataggattt atttatgaaa cctaatgaag caggattaaa tactgtatta aaataggttc 10800 gctgtctttt aaacaaatgg agatgatgat tactaagtca cattgacttt aatatgaggt 10860 atcactatac cttaacatat ttgttaaaac gtatactgta tacattttgt gtattttaat 10920 acttaatact atgaaaacaa gtaattgtaa acgtatcttg tcagattaca ataggaatga 10980 acatattggt gacatcgagt taaagtttat atttccccta aatatgctgc gattccaata 11040 tattcatgta ggttttcaag cagaaataaa ccttgtaaca agttactgac taaacagcct 11100 gacaagtatg tatatatgtt taaaattcaa taaataaaga cccagtcttc taaattataa 11160 aaatttaaat tagtcttgca caaattaaat tattcatcac aaaagatgta ttgttatttt 11220 taagtcattt aagccctaaa tccctaaaga ttagatacaa attttttttt gccagagtat 11280 aaattgtcag aatttatttt taaatatatt ttttaaaact accagtaaga aattttaaat 11340 taaacccatt tgttaaagga tatagtgccc aagttatacg gtgacctacc tttgtcaata 11400 tttagcatta tgtatttcaa attatccaat atacatgtca tatatatttt tatatgttgc 11460 atatataaaa gatatacacg atttatgtga atcctatgta aatattttgt tccagaaaag 11520 tacaaaataa taaaggtaaa aataatccat aattttcagg accacagact aagctgtcaa 11580 aattaacatt gatttttttt agggccagaa taccaaaata gctcctctct tccccaaaaa 11640 ttggacaatt tcaaatgcaa aataattcat tatttaatat atgagttgct tccactattt 11700 ggtttccttt aaaaaaaaaa aaaaaacact ctcataggac atgtttcatt ttgttccttt 11760 taggagtagt aaattagatt ttttccccat acaaagcttt cttttaccag aaagatac tt 11820 ctggcagaag aggagaaagg agctcttttc tttatggttc acacgactgt ctcctgtcct 11880 cactactttg cttaaagtgc tcaaattcca ccactaactc acagttgtct aatctaagca 11940 aagtctaatc ccctttgatc tctgagtcta ccttcccttt catctctcta ctacttacta 12000 atgtgaatat ctttttttca aacttgacct tcattttgct ttcacaatac tatactctcc 12060 atggattttc ccttacttga atccatcttt ataaccctat tcctttctca tatttagtac 12120 tgtgggccaa tggacaacct ttaatcatct tttctacact gactctcaga tattctatct 12180 gctctcacgg actcatctat gtaccatgaa taaagttcca aaatctacgt attcatccta 12240 agtctctttc cagttcccct tcttacattg cctatctgcc atttctccct ttaataacct 12300 atacttcact taaattcaac atatcaaaaa taacaggccg ggcacggtgg ctcatgactg 12360 taatcccagc actttgggag gccgaggcgg gtggatcacc tgaggtcagg agtttcagac 12420 cagcctgacc aacagggtga aaccccgtct ccacttaaaa tacaaaaatt agccaggtgt 12480 ggtggcatgt gcctatagtc ccagttactc aagaggctga gacaggagaa ttgcttgaac 12540 ccaggaggcg gaggttgcag tgagctgaga tcacaccact gcactccatc ctgggtgaca 12600 gaacaagact gactcaaaaa aataaataat tcaccagccc ctgactgcta ctgctatctc 12660 tttctaccca actttccctc ctttcgactc tgtcacacca tctttctcac ttctttatat 12720 ccattaatat gaccagcatg ttcccagtca cagaagcctg aaacctggaa gacatctcta 12780 gcttttccct caacttcgta aactacttct tttgtataag ccaccaagtt caatacaatc 12840 ttctcttgaa acatctctta atcttataag ctttcttccc caaagtctgt ctttaacttg 12900 agtgctagat tatattagtc tcctctttcc tcctaatttt atgtccctct ccctgtcaat 12960cttctgaaaa gaaatttgta atggctgcat attgcttgga 13000 <210> 4 <211> 25 <212> DNA <213> Artificial sequence <220> <223> Primer <400> 4 ccgtggaaga ccaatataaa caatt 25 <210> 5 <211> 28 <212> DNA <213> Artificial sequence <220> <223> Primer <400> 5 agtccttctg agctgatttt ctatttct 28 <210> 6 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Probe <400> 6 aaccaacagc atagtcaaat a 21 <210> 7 <211> 23 <212> DNA <213> Artificial sequence <220> <223> Primer <400> 7 cttcaatgaa acgtgggaga act 23 <210> 8 <211> 22 <212> DNA <213> Artificial sequence <220> <223> Primer <400> 8 tctctaggcc caaccaaaat tc 22 <210> 9 <211> 24 <212> DNA <213> Artificial sequence <220> <223> Probe <400> 9 aaatatggtt ttgggaggct tgat 24 <210> 10 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Primer <400> 10 aaattttagc caatggcctc c 21 <210> 11 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Primer <400> 11 tgtcattaat ttggcccttc g 21 <210> 12 <211> 32 <212> DNA <213> Artificial sequence <220> <223> Probe <400> 12 tcagttggga catggtctta aagactttgt cc 32 <210> 13 <211> 27 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 13 gcgtttgctc ttcttcttgc gtttttt 27 <210> 14 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 14 tgaattaatg tccatggact 20 <210> 15 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 15 agttcttggt gctcttggct 20 <210> 16 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 16 gttgtctttc cagtcttcca 20 <210> 17 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 17 gctttgtgat cccaagtaga 20 <210> 18 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 18 ggttgttttc tccacactca 20 <210> 19 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 19 accttccatt ttgagacttc 20 <210> 20 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 20 tacacatact ctgtgctgac 20 <210> 21 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 21 tttcagcatg tatctcttaa 20 <210> 22 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 22 accaggttaa tcttcctaat 20 <210> 23 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 23 agttatgatc attgtaagcc 20 <210> 24 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 24 ggagtggttc ttttcacagc 20 <210> 25 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 25 tggtcctttt aacttccaat 20 <210> 26 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 26 gcattgagtt aactaattca 20 <210> 27 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 27 catccatata cagtcaagag 20 <210> 28 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 28 cacttgtatg ttcacctctg 20 <210> 29 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 29 tctattcgat gttgaattaa 20 <210> 30 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 30 ttgattttca atttcaagca 20 <210> 31 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 31 ggctctggag atagagaatc 20 <210> 32 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 32 tcttgatttt ggctctggag 20 <210> 33 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 33 gtcattaatt tggcccttcg 20 <210> 34 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 34 ttcactggtt tgcagcgata 20 <210> 35 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 35 cttagatttt gctcttggtt 20 <210> 36 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 36 gcttagattt tgctcttggt 20 <210> 37 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 37 ggcttagatt ttgctcttgg 20 <210> 38 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 38 gctcatatga tgccttttaa 20 <210> 39 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 39 agctcatatg atgcctttta 20 <210> 40 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 40 tagctcatat gatgcctttt 20 <210> 41 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 41 tattagctca tatgatgcct 20 <210> 42 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 42 ccacaaatat gttcattctt 20 <210> 43 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 43 tgccacaaat atgttcattc 20 <210> 44 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 44 taaactttaa ctcgatgcca 20 <210> 45 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 45 acgaggttta tttctacttg 20 <210> 46 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 46 catgcttgtc aggctgttta 20 <210> 47 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 47 tacatgcttg tcaggctgtt 20 <210> 48 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 48 tccaactcag aaattcattg 20 <210> 49 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 49 ttgctgggta ttactctttt 20 <210> 50 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 50 tttgcttgct gggtattact 20 <210> 51 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 51 tggcatgaat gatatttcct 20 <210> 52 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 52 gcaataccat gatcacatct 20 <210> 53 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 53 actccgtaaa tatttcagca 20 <210> 54 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 54 tggcaaatgt agcatttttt 20 <210> 55 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 55 aagccataca ttctatatat 20 <210> 56 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 56 tcagctattt attaacatcc 20 <210> 57 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 57 gaggaatatt tgcaagtctt 20 <210> 58 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 58 ttggtagagg aatatttgca 20 <210> 59 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 59 accagtgaaa tgtgttacat 20 <210> 60 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 60 tgaaagtctg attatccttt 20 <210> 61 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 61 atttaatggg tacagactcg 20 <210> 62 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 62 ataccatgat cacatctaag 20 <210> 63 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 63 gacctttaac tccgtaaata 20 <210> 64 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 64 tctgagtcc agtgaccttt 20 <210> 65 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 65 atgtaagaat catcttacca 20 <210> 66 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 66 agcatttttc tagatggcg 20 <210> 67 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 67 cattttatga agttgtggca 20 <210> 68 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 68 cacttgacat tttatgaagt 20 <210> 69 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 69 tcctggtcct tttaacttcc 20 <210> 70 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 70 gacatcattc atggattttt 20 <210> 71 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 71 agtacttaga catcattcat 20 <210> 72 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 72 ggtcaaatag tactgagcat 20 <210> 73 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 73 ggtagtttga atacaaaatt 20 <210> 74 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 74 ctctgttata aatggtggta 20 <210> 75 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 75 ttcttggtgc tcttggcttg 20 <210> 76 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 76 ggtacattca gcaggaatgc 20 <210> 77 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 77 ggacattgcc agtaatcgca 20 <210> 78 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 78 cactcatcat gccaccacca 20 <210> 79 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 79 ctccacactc atcatgccac 20 <210> 80 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 80 tttctccaca ctcatcatgc 20 <210> 81 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 81 gctgacgatt tttagttttg 20 <210> 82 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 82 cacatactct gtgctgacga 20 <210> 83 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 83 tttacacata ctctgtgctg 20 <210> 84 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 84 gctttcttac cagaattaag 20 <210> 85 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 85 gttgctttct taccagaatt 20 <210> 86 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 86 ataccagatt attagaccac 20 <210> 87 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 87 gcttcatttt aaggattgct 20 <210> 88 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 88 gcttaatgta tgctttcttt 20 <210> 89 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 89 gttaagtatt cctagagact 20 <210> 90 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 90 tccatatttg tatttctctg 20 <210> 91 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 91 cgtaagttta tcttcctttc 20 <210> 92 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 92 cacattaact tggaatgagg 20 <210> 93 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 93 ccagattatt agaccacatt 20 <210> 94 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 94 taccagatta ttagaccaca 20 <210> 95 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 95 gctttctctt aaggatttaa 20 <210> 96 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 96 ctcaagcttt ctcttaagga 20 <210> 97 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 97 tctcaagctt tctcttaagg 20 <210> 98 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 98 ccatttgttt aaagacagcg 20 <210> 99 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 99 acttagtagt catctccatt 20 <210> 100 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 100 gacttagtag tcatctccat 20 <210> 101 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 101 tgacttagta gtcatctcca 20 <210> 102 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 102 gtcaatgtga cttagtagtc 20 <210> 103 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 103 accatataac ttgggcacta 20 <210> 104 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 104 ggtcaccata taacttgggc 20 <210> 105 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 105 gacaaaggta ggtcaccata 20 <210> 106 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 106 gccaatatga gtgaaatgga 20 <210> 107 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 107 tttctaaaca agtgcttccc 20 <210> 108 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 108 ctgcatatta gaatacatgg 20 <210> 109 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 109 cataatttga accaggcttt 20 <210> 110 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 110 aaatcttgat tttggc 16 <210> 111 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 111 ttcttggtgc tcttgg 16 <210> 112 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 112 cgtggaactg ttttct 16 <210> 113 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 113 cttgatcaat tctgga 16 <210> 114 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 114 gtcttgatca attctg 16 <210> 115 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 115 tggctctgga gataga 16 <210> 116 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 116 tgattttggc tctgga 16 <210> 117 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 117 cttgattttg gctctg 16 <210> 118 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 118 ggtgctcttg gcttgg 16 <210> 119 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 119 atcttctcta ggccca 16 <210> 120 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 120 gttgtctttc cagtct 16 <210> 121 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 121 gtttgttgtc tttcca 16 <210> 122 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 122 gtgatttccc aagtaa 16 <210> 123 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 123 tggatcaaca ttttgg 16 <210> 124 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 124 gaatctgttg gatgga 16 <210> 125 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 125 ttctgaatct gttgga 16 <210> 126 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 126 gttcattcaa agcttt 16 <210> 127 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 127 cagttcattc aaagct 16 <210> 128 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 128 ttaacttgga atgagg 16 <210> 129 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 129 agaccacatt aacttg 16 <210> 130 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 130 agattattag accaca 16 <210> 131 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 131 ccagattatt agacca 16 <210> 132 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 132 atctatttct caagct 16 <210> 133 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 133 atagacagtg acttta 16 <210> 134 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 134 atactctgtg ctgacg 16 <210> 135 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 135 actctgtgct gacgat 16 <210> 136 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 136 acatactctg tgctga 16 <210> 137 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 137 gtagcaaaat gaagca 16 <210> 138 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 138 tcaaacattt actcca 16 <210> 139 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 139 taaagacagc gaactt 16 <210> 140 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 140 tgtttaaaga cagcga 16 <210> 141 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 141 agtagtcatc tccatt 16 <210> 142 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 142 ttagtagtca tctcca 16 <210> 143 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 143 acaaaggtag gtcacc 16 <210> 144 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 144 tgacatgtat attgga 16 <210> 145 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 145 gtacttttct ggaaca 16 <210> 146 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 146 cttggtctaa agagcc 16 <210> 147 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 147 tctgtagatg ttggca 16 <210> 148 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 148 ataatttgaa ccaggc 16 <210> 149 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 149 agcagacatt ggtcgg 16 <210> 150 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 150 cagatgtac agatta 16 <210> 151 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 151 gtagaggaat atttgc 16 <210> 152 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 152 aaagtctgat tatcct 16 <210> 153 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 153 tagtcttgaa tccctt 16 <210> 154 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 154 aagttgtcat tgcaga 16 <210> 155 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 155 tctgtataag tcccaa 16 <210> 156 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 156 gatgcatgta tcccag 16 <210> 157 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 157 gtaagtttat cttcct 16 <210> 158 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 158 ctgttactgt atttcc 16 <210> 159 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 159 tgcagcgata gatcat 16 <210> 160 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 160 ctggtttgca gcgata 16 <210> 161 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 161 ctttgatttc actggt 16 <210> 162 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 162 gttcttctca gttcct 16 <210> 163 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 163 gttagttagt tgctct 16 <210> 164 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 164 gtatgttcac ctctgt 16 <210> 165 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 165 gagttgctgg gtctga 16 <210> 166 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 166 aaacttgaga gttgct 16 <210> 167 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 167 gtccatggac tacctg 16 <210> 168 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 168 ccatctattc gatgtt 16 <210> 169 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 169 tagattttgc tcttgg 16 <210> 170 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 170 ccattttgag acttcc 16 <210> 171 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 171 tagagtataa ccttcc 16 <210> 172 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 172 ctcatcatgc caccac 16 <210> 173 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 173 cacactcatc atgcca 16 <210> 174 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 174 gagaaatgta aacggt 16 <210> 175 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 175 ctcatatgat gccttt 16 <210> 176 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 176 agctcatatg atgcct 16 <210> 177 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 177 ttagctcata tgatgc 16 <210> 178 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 178 ttgtgatatt agctca 16 <210> 179 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 179 actgggaaag ttgtga 16 <210> 180 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 180 aatctgacaa gattcc 16 <210> 181 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 181 ctcgatgcca caaata 16 <210> 182 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 182 taactcgatg ccacaa 16 <210> 183 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 183 tgcttgtcag gctgtt 16 <210> 184 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 184 acaggcttgt caggct 16 <210> 185 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 185 cagttatgct gatgtg 16 <210> 186 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 186 aagttttaga gccatg 16 <210> 187 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 187 ctgtcacttc accaga 16 <210> 188 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 188 tggattctga catagc 16 <210> 189 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 189 aaccttttag cagcac 16 <210> 190 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 190 ttgcttgctg ggtatt 16 <210> 191 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 191 gtcttttata gggtgt 16 <210> 192 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 192 ttgtcttagg tatgtc 16 <210> 193 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 193 ctctgattgt cttagg 16 <210> 194 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 194 gtatttctct gattgt 16 <210> 195 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 195 tttttcttag atggcg 16 <210> 196 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 196 gggctttcac tggtag 16 <210> 197 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 197 acagtcaaga gatgtg 16 <210> 198 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 198 atcttgattt tggctctgga 20 <210> 199 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 199 gttcttggtg ctcttggctt 20 <210> 200 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 200 tagttcttgg tgctcttggc 20 <210> 201 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 201 aggttgtttt ctccacactc 20 <210> 202 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 202 acacatactc tgtgctgacg 20 <210> 203 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 203 gtggcaaatg tagcattttt 20 <210> 204 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 204 gccacaaata tgttcattct 20 <210> 205 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 205 tcttggtgct cttggct 17 <210> 206 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 206 ccagattatt agaccac 17 <210> 207 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 207 gctcatatga tgccttt 17 <210> 208 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 208 gcttgtcagg ctgttta 17 <210> 209 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 209 tgtcttgatc aattct 16 <210> 210 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 210 ttgattttgg ctctgg 16 <210> 211 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 211 actggtttgc agcgat 16 <210> 212 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 212 agattttgct cttggt 16 <210> 213 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 213 cattttgaga cttcca 16 <210> 214 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 214 tccattttga gacttc 16 <210> 215 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 215 accagattat tagacc 16 <210> 216 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 216 tagacagtga ctttaa 16 <210> 217 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 217 tgagaaatgt aaacgg 16 <210> 218 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 218 gctcatatga tgcctt 16 <210> 219 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 219 cttagtagtc atctcc 16 <210> 220 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 220 agagtataac cttcca 16 <210> 221 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 221 cagattatta gaccac 16 <210> 222 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 222 tagctcatat gatgcc 16 <210> 223 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 223 ccttttagca gcacta 16 <210> 224 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 224 accttttagc agcact 16 <210> 225 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 225 aaacctttta gcagca 16 <210> 226 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 226 atctgtagat gttggc 16 <210> 227 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 227 taatttgaac caggct 16 <210> 228 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 228 ttttcttaga tggcgt 16 <210> 229 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 229 atctgtataa gtccca 16 <210> 230 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 230 aatctgtata agtccc 16 <210> 231 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 231 atgcatgtat cccagt 16 <210> 232 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 232 tagatgcatg tatccc 16 <210> 233 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 233 agtagttctt ggtgctcttg 20 <210> 234 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 234 ggttcttgaa tactagtcct 20 <210> 235 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 235 aaatttctgt gggttcttga 20 <210> 236 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 236 tggcttggaa gatagagaaa 20 <210> 237 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 237 gtattagaat cacagcatat 20 <210> 238 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 238 gattttcaat ttcaagcaac 20 <210> 239 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 239 agcttaattg tgaacatttt 20 <210> 240 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 240 gaaggagctt aattgtgaac 20 <210> 241 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 241 caataaaaag aaggagctta 20 <210> 242 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 242 gaacaataaa aagaaggagc 20 <210> 243 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 243 ctggaggaaa taactagagg 20 <210> 244 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 244 attctggagg aaataactag 20 <210> 245 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 245 tcaaatgatg aattgtcttg 20 <210> 246 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 246 ttgattttgg ctctggagat 20 <210> 247 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 247 gcaaatcttg attttggctc 20 <210> 248 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 248 atagcaaatc ttgattttgg 20 <210> 249 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 249 cgtctaacat agcaaatctt 20 <210> 250 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 250 tggctaaaat ttttacatcg 20 <210> 251 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 251 ccattggcta aaatttttac 20 <210> 252 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 252 aggaggccat tggctaaaat 20 <210> 253 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 253 tgaaggaggc cattggctaa 20 <210> 254 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 254 gtcccaactg aaggaggcca 20 <210> 255 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 255 ccatgtccca actgaaggag 20 <210> 256 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 256 agaccatgtc ccaactgaag 20 <210> 257 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 257 tttaagacca tgtcccaact 20 <210> 258 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 258 gtctttaaga ccatgtccca 20 <210> 259 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 259 ggacaaagtc tttaagacca 20 <210> 260 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 260 tcttatggac aaagtcttta 20 <210> 261 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 261 tatgtcatta atttggccct 20 <210> 262 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 262 gatcaaatat gttgagtttt 20 <210> 263 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 263 gactgatcaa atatgttgag 20 <210> 264 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 264 tcttctttga tttcactggt 20 <210> 265 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 265 cttctcagtt ccttttcttc 20 <210> 266 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 266 gttcttctca gttccttttc 20 <210> 267 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 267 tgtagttctt ctcagttcct 20 <210> 268 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 268 tatatgtagt tcttctcagt 20 <210> 269 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 269 gtttatatgt agttcttctc 20 <210> 270 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 270 tgtagtttat atgtagttct 20 <210> 271 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 271 gacttgtagt ttatatgtag 20 <210> 272 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 272 tcatttttga cttgtagttt 20 <210> 273 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 273 cctcttcatt tttgacttgt 20 <210> 274 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 274 tctttacctc ttcatttttg 20 <210> 275 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 275 catattcttt acctcttcat 20 <210> 276 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 276 gtgacatatt ctttacctct 20 <210> 277 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 277 gagttcaagt gacatattct 20 <210> 278 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 278 tgagttgagt tcaagtgaca 20 <210> 279 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 279 agttttgagt tgagttcaag 20 <210> 280 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 280 tcaagttttg agttgagttc 20 <210> 281 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 281 ggaggctttc aagttttgag 20 <210> 282 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 282 tttcttctag gaggctttca 20 <210> 283 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 283 attttttctt ctaggaggct 20 <210> 284 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 284 gtagaatttt ttcttctagg 20 <210> 285 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 285 gctcttctaa atatttcact 20 <210> 286 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 286 agttagttag ttgctcttct 20 <210> 287 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 287 caggttgatt ttgaattaag 20 <210> 288 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 288 tttcaggttg attttgaatt 20 <210> 289 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 289 ggagtttcag gttgattttg 20 <210> 290 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 290 gttctggagt ttcaggttga 20 <210> 291 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 291 ttaagtgaag ttacttctgg 20 <210> 292 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 292 tgctattatc ttgtttttct 20 <210> 293 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 293 ggtctttgat gctattatct 20 <210> 294 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 294 gaaggtcttt gatgctatta 20 <210> 295 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 295 ctggagaagg tctttgatgc 20 <210> 296 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 296 ctgagctgat tttctatttc 20 <210> 297 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 297 atttctgtgg gttcttgaat 20 <210> 298 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 298 gagaaatttc tgtgggttct 20 <210> 299 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 299 gatagagaaa tttctgtggg 20 <210> 300 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 300 cttggaagat agagaaattt 20 <210> 301 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 301 gtgctcttgg cttggaagat 20 <210> 302 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 302 cttggtgctc ttggcttgga 20 <210> 303 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 303 gggagtagtt cttggtgctc 20 <210> 304 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 304 ctgaagaaag ggagtagttc 20 <210> 305 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 305 atcatgtttt acatttctta 20 <210> 306 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 306 gccatcatgt tttacatttc 20 <210> 307 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 307 gaatgccatc atgttttaca 20 <210> 308 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 308 caggaatgcc atcatgtttt 20 <210> 309 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 309 gacttcttaa ctctatatat 20 <210> 310 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 310 ctagacttct taactctata 20 <210> 311 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 311 gacctagact tcttaactct 20 <210> 312 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 312 ggaagcagac ctagacttct 20 <210> 313 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 313 tctggaagca gacctagact 20 <210> 314 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 314 tcttctggaa gcagacctag 20 <210> 315 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 315 ctaatcttta gggatttagg 20 <210> 316 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 316 tgtatctaat ctttagggat 20 <210> 317 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 317 taacttgggc actatatcct 20 <210> 318 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 318 attgacaaag gtaggtcacc 20 <210> 319 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 319 atatgacatg tatattggat 20 <210> 320 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 320 ttttgtactt ttctggaaca 20 <210> 321 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 321 tagtctgtgg tcctgaaaat 20 <210> 322 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 322 agcttagtct gtggtcctga 20 <210> 323 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 323 gacagcttag tctgtggtcc 20 <210> 324 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 324 gtattctggc cctaaaaaaa 20 <210> 325 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 325 attttggtat tctggcccta 20 <210> 326 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 326 tttgcatttg aaattgtcca 20 <210> 327 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 327 ggaagcaact catatattaa 20 <210> 328 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 328 tatcagaaaa agatacctga 20 <210> 329 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 329 ataatagcta ataatgtggg 20 <210> 330 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 330 tgcagataat agctaataat 20 <210> 331 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 331 tgtcattgca gataatagct 20 <210> 332 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 332 taaaagttgt cattgcagat 20 <210> 333 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 333 cggattttta aaagttgtca 20 <210> 334 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 334 gggattcgga tttttaaaag 20 <210> 335 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 335 tttgggattc ggatttttaa 20 <210> 336 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 336 acgcttattt gggattcgga 20 <210> 337 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 337 tttaagagat ttacaagtca 20 <210> 338 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 338 gactacctgt ttttaaaagc 20 <210> 339 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 339 tatggtgact acctgttttt 20 <210> 340 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 340 actttgctgt attataaact 20 <210> 341 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 341 attgtattta actttgctgt 20 <210> 342 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 342 gagcaactaa cttaataggt 20 <210> 343 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 343 gaaatgagca actaacttaa 20 <210> 344 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 344 aatcaaagaa atgagcaact 20 <210> 345 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 345 accttcttcc acctgagtt 20 <210> 346 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 346 cacgaatgta accttcttcc 20 <210> 347 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 347 ttaacttgca cgaatgtaac 20 <210> 348 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 348 tatatatacc aatatttgcc 20 <210> 349 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 349 tcttaactct atatatacca 20 <210> 350 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 350 ctttaagtga agttacttct 20 <210> 351 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 351 tctacttact ttaagtgaag 20 <210> 352 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 352 gaaccctctt tattttctac 20 <210> 353 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 353 acataaacat gaaccctctt 20 <210> 354 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 354 ccacattgaa aacataaaca 20 <210> 355 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 355 gcatgcctta gaaatatttt 20 <210> 356 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 356 caatgcaaca aagtatttca 20 <210> 357 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 357 ctggagatta tttttcttgg 20 <210> 358 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 358 ttcatatata acattaggga 20 <210> 359 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 359 tcagtgtttt catataac 20 <210> 360 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 360 gacatagtgt tctagattgt 20 <210> 361 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 361 caatagtgta atgacatagt 20 <210> 362 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 362 ttacttacct tcagtaattt 20 <210> 363 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 363 atcttttcca tttactgtat 20 <210> 364 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 364 agaaaaagcc cagcatattt 20 <210> 365 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 365 gtatgcttct ttcaaatagc 20 <210> 366 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 366 ccttcccctt gtatgcttct 20 <210> 367 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 367 cctgtaacac tatcataatc 20 <210> 368 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 368 tgacttacct gattttctat 20 <210> 369 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 369 gatgggacat accattaaaa 20 <210> 370 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 370 gtgaaagatg ggacatacca 20 <210> 371 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 371 cctgtgtgaa agatgggaca 20 <210> 372 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 372 cattggctgc tatgaattaa 20 <210> 373 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 373 gatgacattg gctgctatga 20 <210> 374 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 374 gagaaacatg atctaatttg 20 <210> 375 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 375 atggaaagct attgtgtggt 20 <210> 376 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 376 gtctaaagag ccaatatgag 20 <210> 377 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 377 aatcttggtc taaagagcca 20 <210> 378 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 378 gagatttaca agtcaaaaat 20 <210> 379 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 379 atttaacttt gctgtattat 20 <210> 380 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 380 atcaatgcta aatgaaatca 20 <210> 381 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 381 tattttctgg agattatttt 20 <210> 382 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 382 aaaatgaata ttggcaattc 20 <210> 383 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 383 aaagtcaatg tgacttagta 20 <210> 384 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 384 aaggtatagt gatacctcat 20 <210> 385 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 385 cagcaggaat gccatcatgt 20 <210> 386 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 386 tgatggcata catgccactt 20 <210> 387 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 387 tgctgggtct gatggcatac 20 <210> 388 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 388 gagttgctgg gtctgatggc 20 <210> 389 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 389 aaaacttgag agttgctggg 20 <210> 390 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 390 gacatgaaaa acttgagagt 20 <210> 391 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 391 acatcacagt agacatgaaa 20 <210> 392 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 392 agttttgtga tccatctatt 20 <210> 393 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 393 tgaagttttg tgatccatct 20 <210> 394 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 394 cgtttcattg aagttttgtg 20 <210> 395 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 395 ccatatttgt agttctccca 20 <210> 396 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 396 aaaccatatt tgtagttctc 20 <210> 397 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 397 aattctccat caagcctccc 20 <210> 398 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 398 atcttctcta ggcccaacca 20 <210> 399 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 399 gagtatatct tctctaggcc 20 <210> 400 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 400 ctatggagta tatcttctct 20 <210> 401 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 401 gcttcactat ggagtatatc 20 <210> 402 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 402 agattgcttc actatggagt 20 <210> 403 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 403 ccagtcttcc aactcaattc 20 <210> 404 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 404 gtctttccag tcttccaact 20 <210> 405 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 405 gtttgttgtc tttccagtct 20 <210> 406 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 406 aatgtttgtt gtctttccag 20 <210> 407 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 407 cgtgatttcc caagtaaaaa 20 <210> 408 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 408 gttttccggg attgcattgg 20 <210> 409 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 409 tctttgtttt ccgggattgc 20 <210> 410 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 410 ccaaatcttt gttttccggg 20 <210> 411 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 411 acaccaaatc tttgttttcc 20 <210> 412 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 412 agaaaacacc aaatctttgt 20 <210> 413 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 413 caagtagaaa acaccaaatc 20 <210> 414 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 414 gatcccaagt agaaaacacc 20 <210> 415 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 415 tttgctttgt gatcccaagt 20 <210> 416 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 416 tccttttgct ttgtgatccc 20 <210> 417 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 417 gaagtgtcct tttgctttgt 20 <210> 418 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 418 tgccaccacc agcctcctga 20 <210> 419 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 419 ctcatcatgc caccaccagc 20 <210> 420 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 420 ccatttaggt tgttttctcc 20 <210> 421 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 421 atatttacca tttaggttgt 20 <210> 422 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 422 cttggtttgt tatatttacc 20 <210> 423 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 423 atagagtata accttccatt 20 <210> 424 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 424 ttttatagag tataaccttc 20 <210> 425 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 425 tggttgattt tatagagtat 20 <210> 426 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 426 attttggttg attttataga 20 <210> 427 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 427 tcaacatttt ggttgatttt 20 <210> 428 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 428 ggatggatca acattttggt 20 <210> 429 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 429 gttggatgga tcaacatttt 20 <210> 430 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 430 tctgttggat ggatcaacat 20 <210> 431 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 431 ctgaatctgt tggatggatc 20 <210> 432 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 432 agctttctga atctgttgga 20 <210> 433 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 433 cattcaaagc tttctgaatc 20 <210> 434 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 434 gttcattcaa agctttctga 20 <210> 435 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 435 tcagttcatt caaagctttc 20 <210> 436 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 436 gcctcagttc attcaaagct 20 <210> 437 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 437 atttgcctca gttcattcaa 20 <210> 438 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 438 ttaaatttgc ctcagttcat 20 <210> 439 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 439 gccttttaaa tttgcctcag 20 <210> 440 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 440 aggatttaat accagattat 20 <210> 441 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 441 cttaaggatt taataccaga 20 <210> 442 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 442 tctcttaagg atttaatacc 20 <210> 443 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 443 gacagtgact ttaagataaa 20 <210> 444 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 444 tgtgattgta tgtttaatct 20 <210> 445 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 445 aggttatgtg attgtatgtt 20 <210> 446 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 446 ctttaaggtt atgtgattgt 20 <210> 447 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 447 ggtattcttt aaggttatgt 20 <210> 448 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 448 attgattccc acatcacaaa 20 <210> 449 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 449 ctaaaattga ttcccacatc 20 <210> 450 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 450 ccatctaaaa ttgattccca 20 <210> 451 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 451 ttgtgatatt agctcatatg 20 <210> 452 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 452 actagttttt taaactggga 20 <210> 453 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 453 gtcaagttta gagttttaac 20 <210> 454 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 454 tatttagtca agtttagagt 20 <210> 455 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 455 gatttttaca catactctgt 20 <210> 456 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 456 ctgcttcatt aggtttcata 20 <210> 457 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 457 cagcaggaat gccatcatgt 20 <210> 458 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 458 tgatggcata catgccactt 20 <210> 459 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 459 tgctgggtct gatggcatac 20 <210> 460 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 460 gagttgctgg gtctgatggc 20 <210> 461 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 461 aaaacttgag agttgctggg 20 <210> 462 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 462 gacatgaaaa acttgagagt 20 <210> 463 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 463 acatcacagt agacatgaaa 20 <210> 464 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 464 agttttgtga tccatctatt 20 <210> 465 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 465 tgaagttttg tgatccatct 20 <210> 466 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 466 cgtttcattg aagttttgtg 20 <210> 467 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 467 ccatatttgt agttctccca 20 <210> 468 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 468 aaaccatatt tgtagttctc 20 <210> 469 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 469 aattctccat caagcctccc 20 <210> 470 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 470 atcttctcta ggcccaacca 20 <210> 471 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 471 gagtatatct tctctaggcc 20 <210> 472 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 472 ctatggagta tatcttctct 20 <210> 473 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 473 gcttcactat ggagtatatc 20 <210> 474 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 474 agattgcttc actatggagt 20 <210> 475 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 475 ccagtcttcc aactcaattc 20 <210> 476 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 476 gtctttccag tcttccaact 20 <210> 477 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 477 gtttgttgtc tttccagtct 20 <210> 478 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 478 aatgtttgtt gtctttccag 20 <210> 479 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 479 cgtgatttcc caagtaaaaa 20 <210> 480 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 480 gttttccggg attgcattgg 20 <210> 481 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 481 tctttgtttt ccgggattgc 20 <210> 482 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 482 ccaaatcttt gttttccggg 20 <210> 483 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 483 acaccaaatc tttgttttcc 20 <210> 484 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 484 agaaaacacc aaatctttgt 20 <210> 485 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 485 caagtagaaa acaccaaatc 20 <210> 486 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 486 gatcccaagt agaaaacacc 20 <210> 487 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 487 tttgctttgt gatcccaagt 20 <210> 488 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 488 tccttttgct ttgtgatccc 20 <210> 489 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 489 gaagtgtcct tttgctttgt 20 <210> 490 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 490 tgccaccacc agcctcctga 20 <210> 491 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 491 ctcatcatgc caccaccagc 20 <210> 492 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 492 ccatttaggt tgttttctcc 20 <210> 493 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 493 atatttacca tttaggttgt 20 <210> 494 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 494 cttggtttgt tatatttacc 20 <210> 495 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 495 atagagtata accttccatt 20 <210> 496 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 496 ttttatagag tataaccttc 20 <210> 497 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 497 tggttgattt tatagagtat 20 <210> 498 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 498 attttggttg attttataga 20 <210> 499 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 499 tcaacatttt ggttgatttt 20 <210> 500 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 500 ggatggatca acattttggt 20 <210> 501 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 501 gttggatgga tcaacatttt 20 <210> 502 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 502 tctgttggat ggatcaacat 20 <210> 503 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 503 ctgaatctgt tggatggatc 20 <210> 504 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 504 agctttctga atctgttgga 20 <210> 505 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 505 cattcaaagc tttctgaatc 20 <210> 506 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 506 gttcattcaa agctttctga 20 <210> 507 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 507 tcagttcatt caaagctttc 20 <210> 508 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 508 gcctcagttc attcaaagct 20 <210> 509 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 509 atttgcctca gttcattcaa 20 <210> 510 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 510 ttaaatttgc ctcagttcat 20 <210> 511 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 511 gccttttaaa tttgcctcag 20 <210> 512 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 512 aggatttaat accagattat 20 <210> 513 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 513 cttaaggatt taataccaga 20 <210> 514 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 514 tctcttaagg atttaatacc 20 <210> 515 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 515 gacagtgact ttaagataaa 20 <210> 516 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 516 tgtgattgta tgtttaatct 20 <210> 517 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 517 aggttatgtg attgtatgtt 20 <210> 518 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 518 ctttaaggtt atgtgattgt 20 <210> 519 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 519 ggtattcttt aaggttatgt 20 <210> 520 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 520 attgattccc acatcacaaa 20 <210> 521 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 521 ctaaaattga ttcccacatc 20 <210> 522 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 522 ccatctaaaa ttgattccca 20 <210> 523 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 523 ttgtgatatt agctcatatg 20 <210> 524 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 524 actagttttt taaactggga 20 <210> 525 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 525 gtcaagttta gagttttaac 20 <210> 526 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 526 tatttagtca agtttagagt 20 <210> 527 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 527 gatttttaca catactctgt 20 <210> 528 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 528 ctgcttcatt aggtttcata 20 <210> 529 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 529 cagcaggaat gccatcatgt 20 <210> 530 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 530 tgatggcata catgccactt 20 <210> 531 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 531 tgctgggtct gatggcatac 20 <210> 532 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 532 gagttgctgg gtctgatggc 20 <210> 533 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 533 aaaacttgag agttgctggg 20 <210> 534 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 534 gacatgaaaa acttgagagt 20 <210> 535 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 535 acatcacagt agacatgaaa 20 <210> 536 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 536 agttttgtga tccatctatt 20 <210> 537 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 537 tgaagttttg tgatccatct 20 <210> 538 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 538 cgtttcattg aagttttgtg 20 <210> 539 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 539 ccatatttgt agttctccca 20 <210> 540 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 540 aaaccatatt tgtagttctc 20 <210> 541 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 541 aattctccat caagcctccc 20 <210> 542 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 542 atcttctcta ggcccaacca 20 <210> 543 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 543 gagtatatct tctctaggcc 20 <210> 544 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 544 ctatggagta tatcttctct 20 <210> 545 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 545 gcttcactat ggagtatatc 20 <210> 546 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 546 agattgcttc actatggagt 20 <210> 547 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 547 ccagtcttcc aactcaattc 20 <210> 548 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 548 gtctttccag tcttccaact 20 <210> 549 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 549 gtttgttgtc tttccagtct 20 <210> 550 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 550 aatgtttgtt gtctttccag 20 <210> 551 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 551 cgtgatttcc caagtaaaaa 20 <210> 552 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 552 gttttccggg attgcattgg 20 <210> 553 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 553 tctttgtttt ccgggattgc 20 <210> 554 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 554 ccaaatcttt gttttccggg 20 <210> 555 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 555 acaccaaatc tttgttttcc 20 <210> 556 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 556 agaaaacacc aaatctttgt 20 <210> 557 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 557 caagtagaaa acaccaaatc 20 <210> 558 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 558 gatcccaagt agaaaacacc 20 <210> 559 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 559 tttgctttgt gatcccaagt 20 <210> 560 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 560 tccttttgct ttgtgatccc 20 <210> 561 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 561 gaagtgtcct tttgctttgt 20 <210> 562 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 562 tgccaccacc agcctcctga 20 <210> 563 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 563 ctcatcatgc caccaccagc 20 <210> 564 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 564 ccatttaggt tgttttctcc 20 <210> 565 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 565 atatttacca tttaggttgt 20 <210> 566 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 566 cttggtttgt tatatttacc 20 <210> 567 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 567 atagagtata accttccatt 20 <210> 568 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 568 ttttatagag tataaccttc 20 <210> 569 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 569 tggttgattt tatagagtat 20 <210> 570 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 570 attttggttg attttataga 20 <210> 571 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 571 tcaacatttt ggttgatttt 20 <210> 572 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 572 ggatggatca acattttggt 20 <210> 573 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 573 gttggatgga tcaacatttt 20 <210> 574 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 574 tctgttggat ggatcaacat 20 <210> 575 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 575 ctgaatctgt tggatggatc 20 <210> 576 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 576 agctttctga atctgttgga 20 <210> 577 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 577 cattcaaagc tttctgaatc 20 <210> 578 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 578 gttcattcaa agctttctga 20 <210> 579 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 579 tcagttcatt caaagctttc 20 <210> 580 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 580 gcctcagttc attcaaagct 20 <210> 581 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 581 atttgcctca gttcattcaa 20 <210> 582 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 582 ttaaatttgc ctcagttcat 20 <210> 583 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 583 gccttttaaa tttgcctcag 20 <210> 584 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 584 aggatttaat accagattat 20 <210> 585 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 585 cttaaggatt taataccaga 20 <210> 586 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 586 tctcttaagg atttaatacc 20 <210> 587 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 587 gacagtgact ttaagataaa 20 <210> 588 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 588 tgtgattgta tgtttaatct 20 <210> 589 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 589 aggttatgtg attgtatgtt 20 <210> 590 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 590 ctttaaggtt atgtgattgt 20 <210> 591 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 591 ggtattcttt aaggttatgt 20 <210> 592 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 592 attgattccc acatcacaaa 20 <210> 593 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 593 ctaaaattga ttcccacatc 20 <210> 594 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 594 ccatctaaaa ttgattccca 20 <210> 595 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 595 ttgtgatatt agctcatatg 20 <210> 596 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 596 actagttttt taaactggga 20 <210> 597 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 597 gtcaagttta gagttttaac 20 <210> 598 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 598 tatttagtca agtttagagt 20 <210> 599 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 599 gatttttaca catactctgt 20 <210> 600 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 600 ctgcttcatt aggtttcata 20 <210> 601 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 601 cagtgttatt cagattgtac 20 <210> 602 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 602 agtgtcttac catcatgttt 20 <210> 603 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 603 caccagcctc ctaaaggaga 20 <210> 604 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 604 gaggaggtga agtcagtgag 20 <210> 605 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 605 tagagtagag gaggtgaagt 20 <210> 606 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 606 tgtttgatgt gtttgaatac 20 <210> 607 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 607 gaaacaacaa gggcaaaggc 20 <210> 608 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 608 tgtttgataa cgaccctaag 20 <210> 609 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 609 tttttggtta agtgaccttg 20 <210> 610 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 610 gtagaagttt tcagggatgg 20 <210> 611 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 611 aggaagtaga agttttcagg 20 <210> 612 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 612 aggtgagtgt gcaggagaaa 20 <210> 613 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 613 ttaaataaag gtgagtgtgc 20 <210> 614 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 614 agtgcaggaa tagaagagat 20 <210> 615 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 615 cattttagtg caggaataga 20 <210> 616 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 616 ctatattctg gagtatatac 20 <210> 617 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 617 cagtattcta tattctggag 20 <210> 618 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 618 gtgccataca gtattctata 20 <210> 619 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 619 ctgtgtgaat atgacattac 20 <210> 620 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 620 tgaggcacac tatttctagt 20 <210> 621 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 621 gacctttaat tatgaggcac 20 <210> 622 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 622 gaatgttgac ctttaattat 20 <210> 623 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 623 ttgttgaatg ttgaccttta 20 <210> 624 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 624 tctactaagt aactatgtga 20 <210> 625 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 625 ctcttttcta ctaagtaact 20 <210> 626 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 626 aaggatctat tgtaaagttt 20 <210> 627 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 627 ctaggacctt atttaaaagg 20 <210> 628 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 628 atttcctagg accttattta 20 <210> 629 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 629 ttgacagtaa gaaaagcaga 20 <210> 630 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 630 ttctcattga cagtaagaaa 20 <210> 631 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 631 agtttttctc attgacagta 20 <210> 632 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 632 attgaatgat agtttttctc 20 <210> 633 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 633 ttgggtttgc aatttattga 20 <210> 634 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 634 agtgtgttgg gtttgcaatt 20 <210> 635 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 635 tattaagtg tgttgggttt 20 <210> 636 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 636 atatattcag tagtttatcg 20 <210> 637 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 637 agatgttggc aggttggcaa 20 <210> 638 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 638 tctgtagatg ttggcaggtt 20 <210> 639 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 639 ttgataattt ttgacctgta 20 <210> 640 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 640 ggctttcttg ataatttgat 20 <210> 641 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 641 gtcttactga tcttcagacc 20 <210> 642 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 642 tttaggtctt actgatcttc 20 <210> 643 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 643 tcagttttag gtcttactga 20 <210> 644 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 644 tgatattctg ttcagatttt 20 <210> 645 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 645 tagagactgc tttgcttaga 20 <210> 646 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 646 aggccaaaag tagagactgc 20 <210> 647 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 647 ggcaaaaaag cagacattgg 20 <210> 648 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 648 aatcagggac atttattaat 20 <210> 649 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 649 tattaatca gggacattat 20 <210> 650 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 650 ctcaaaatat ttaatcaggg 20 <210> 651 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 651 tacctgttct caaaatattt 20 <210> 652 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 652 gtacagatta cctgttctca 20 <210> 653 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 653 ggtgtttgat atttagataa 20 <210> 654 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 654 ttgtctttca gttcataatg 20 <210> 655 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 655 acagtttgtc tttcagttca 20 <210> 656 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 656 tctgagctga taaaagaata 20 <210> 657 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 657 cccaccaaag tgtcttacca 20 <210> 658 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 658 cttcaagaag gaaacccacc 20 <210> 659 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 659 aatagcttca agaaggaaac 20 <210> 660 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 660 acaagtccta agaataggga 20 <210> 661 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 661 gtctagaaca agtcctaaga 20 <210> 662 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 662 tctaataatc aagtccatat 20 <210> 663 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 663 accttctata ttatctaata 20 <210> 664 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 664 gcatgtatct cttaaacagg 20 <210> 665 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 665 gtccagtgac ctttaactcc 20 <210> 666 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 666 tcttaccaaa ctattttctt 20 <210> 667 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 667 gtaatgttta tgttaaagca 20 <210> 668 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 668 ttgtggcaaa tgtagcattt 20 <210> 669 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 669 gagattcac ttgacatttt 20 <210> 670 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 670 ggagcttgag atttcacttg 20 <210> 671 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 671 catcagattt agtaatagga 20 <210> 672 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 672 gttattacat cagattagt 20 <210> 673 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 673 cagcaggaat gcctagaatc 20 <210> 674 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 674 ctccttagac aggttttacc 20 <210> 675 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 675 gtctattctc cttagacagg 20 <210> 676 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 676 atgaatgatt gaatgtagtc 20 <210> 677 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 677 atatgaaggc tgagactgct 20 <210> 678 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 678 ataaattata tgaaggctga 20 <210> 679 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 679 atatttaaga acagacatgt 20 <210> 680 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 680 atttgtaaca gttactactt 20 <210> 681 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 681 cacagcttat ttgtaacagt 20 <210> 682 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 682 gtgactaatg ctaggagtgg 20 <210> 683 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 683 gaatagagtg actaatgcta 20 <210> 684 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 684 atgagagaat agagtgacta 20 <210> 685 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 685 tatactgtat gtctgagttt 20 <210> 686 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 686 aactaattca ttataagcca 20 <210> 687 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 687 tttggatttt aaacatctgt 20 <210> 688 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 688 tgtatgtgct ttttggattt 20 <210> 689 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 689 catggatttt tgtatgtgct 20 <210> 690 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 690 tcattcatgg atttttgtat 20 <210> 691 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 691 acttagacat cattcatgga 20 <210> 692 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 692 gtgagtactt agacatcatt 20 <210> 693 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 693 tttataagtg agtacttaga 20 <210> 694 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 694 gtcttctact ttataagtga 20 <210> 695 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 695 atgaatgtct tctactttat 20 <210> 696 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 696 caaatagtac tgagcattta 20 <210> 697 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 697 ttagaagatt tggagctaca 20 <210> 698 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 698 tcactattag aagatttgga 20 <210> 699 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 699 gggttacact cactattaga 20 <210> 700 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 700 acttacctgt cagcctttta 20 <210> 701 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 701 cttaccagaa ttaagtgagt 20 <210> 702 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 702 aatacaagta caaatgggtt 20 <210> 703 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 703 ctggtaaata caagtacaaa 20 <210> 704 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 704 ggattgctgg taaatacaag 20 <210> 705 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 705 tcattttaag gattgctggt 20 <210> 706 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 706 agttagtagg aagcttcatt 20 <210> 707 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 707 gctattgagt tagtaggaag 20 <210> 708 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 708 agcatggttc ttaataactt 20 <210> 709 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 709 ctttgtagaa aaagacagga 20 <210> 710 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 710 acctggcctt tggtatttgc 20 <210> 711 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 711 agtctttata tggataaact 20 <210> 712 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 712 cgtcattggt agaggaatat 20 <210> 713 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 713 gattatcctt tctataatgc 20 <210> 714 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 714 gtcttgaatc ccttgatcat 20 <210> 715 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 715 ggtgcaacta attgagttgt 20 <210> 716 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 716 gtgtttttta ttggtgcaac 20 <210> 717 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 717 attctcctga aaagaaaagt 20 <210> 718 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 718 atgccaccac caggctccta 20 <210> 719 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 719 atatccttta acaaatgggt 20 <210> 720 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 720 gcactatatc ctttaacaaa 20 <210> 721 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 721 acttgggcac tatatccttt 20 <210> 722 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 722 gaaacatgtc ctatgagagt 20 <210> 723 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 723 ttgagcactt taagcaaagt 20 <210> 724 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 724 ggaatttgag cactttaagc 20 <210> 725 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 725 tagattagac aactgtgagt 20 <210> 726 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 726 aaaatgaagg tcaagtttga 20 <210> 727 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 727 gtgaaagcaa aatgaaggtc 20 <210> 728 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 728 gtattgtgaa agcaaaatga 20 <210> 729 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 729 tggagagtat agtattgtga 20 <210> 730 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 730 aggaatagaa gagataaata 20 <210> 731 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 731 tggagtatat acaaataatg 20 <210> 732 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 732 tgtttacatt gtagattaat 20 <210> 733 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 733 cagaatatat aatatcttgc 20 <210> 734 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 734 tgcaatttat tgaatgatag 20 <210> 735 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 735 cataatacat aatttgaacc 20 <210> 736 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 736 ataattttca gttttaggtc 20 <210> 737 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 737 tttcagtaat gtttatgtta 20 <210> 738 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 738 aatgcctaga atcaataaaa 20 <210> 739 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 739 gtaaatattt gtagattagc 20 <210> 740 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 740 acaaatgtgt aattgtttga 20 <210> 741 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 741 tactaacaaa tgtgtaattg 20 <210> 742 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 742 tgataagtat atttaagaac 20 <210> 743 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 743 ttaacttcca attaattgat 20 <210> 744 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 744 tctgttattt tatcttgctt 20 <210> 745 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 745 atcacaatcc tttttattaa 20 <210> 746 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 746 agagacttga gtaataataa 20 <210> 747 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 747 aacaaaatga aacatgtcct 20 <210> 748 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 748 cagcaggaat gccatcatgt 20 <210> 749 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 749 tgatggcata catgccactt 20 <210> 750 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 750 tgctgggtct gatggcatac 20 <210> 751 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 751 gagttgctgg gtctgatggc 20 <210> 752 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 752 aaaacttgag agttgctggg 20 <210> 753 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 753 gacatgaaaa acttgagagt 20 <210> 754 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 754 acatcacagt agacatgaaa 20 <210> 755 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 755 agttttgtga tccatctatt 20 <210> 756 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 756 tgaagttttg tgatccatct 20 <210> 757 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 757 cgtttcattg aagttttgtg 20 <210> 758 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 758 ccatatttgt agttctccca 20 <210> 759 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 759 aaaccatatt tgtagttctc 20 <210> 760 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 760 aattctccat caagcctccc 20 <210> 761 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 761 atcttctcta ggcccaacca 20 <210> 762 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 762 gagtatatct tctctaggcc 20 <210> 763 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 763 ctatggagta tatcttctct 20 <210> 764 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 764 gcttcactat ggagtatatc 20 <210> 765 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 765 agattgcttc actatggagt 20 <210> 766 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 766 ccagtcttcc aactcaattc 20 <210> 767 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 767 gtctttccag tcttccaact 20 <210> 768 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 768 gtttgttgtc tttccagtct 20 <210> 769 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 769 aatgtttgtt gtctttccag 20 <210> 770 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 770 cgtgatttcc caagtaaaaa 20 <210> 771 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 771 gttttccggg attgcattgg 20 <210> 772 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 772 tctttgtttt ccgggattgc 20 <210> 773 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 773 ccaaatcttt gttttccggg 20 <210> 774 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 774 acaccaaatc tttgttttcc 20 <210> 775 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 775 agaaaacacc aaatctttgt 20 <210> 776 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 776 caagtagaaa acaccaaatc 20 <210> 777 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 777 gatcccaagt agaaaacacc 20 <210> 778 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 778 tttgctttgt gatcccaagt 20 <210> 779 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 779 tccttttgct ttgtgatccc 20 <210> 780 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 780 gaagtgtcct tttgctttgt 20 <210> 781 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 781 tgccaccacc agcctcctga 20 <210> 782 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 782 ctcatcatgc caccaccagc 20 <210> 783 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 783 ccatttaggt tgttttctcc 20 <210> 784 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 784 atatttacca tttaggttgt 20 <210> 785 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 785 cttggtttgt tatatttacc 20 <210> 786 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 786 atagagtata accttccatt 20 <210> 787 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 787 ttttatagag tataaccttc 20 <210> 788 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 788 tggttgattt tatagagtat 20 <210> 789 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 789 attttggttg attttataga 20 <210> 790 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 790 tcaacatttt ggttgatttt 20 <210> 791 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 791 ggatggatca acattttggt 20 <210> 792 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 792 gttggatgga tcaacatttt 20 <210> 793 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 793 tctgttggat ggatcaacat 20 <210> 794 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 794 ctgaatctgt tggatggatc 20 <210> 795 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 795 agctttctga atctgttgga 20 <210> 796 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 796 cattcaaagc tttctgaatc 20 <210> 797 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 797 gttcattcaa agctttctga 20 <210> 798 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 798 tcagttcatt caaagctttc 20 <210> 799 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 799 gcctcagttc attcaaagct 20 <210> 800 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 800 atttgcctca gttcattcaa 20 <210> 801 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 801 ttaaatttgc ctcagttcat 20 <210> 802 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 802 gccttttaaa tttgcctcag 20 <210> 803 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 803 aggatttaat accagattat 20 <210> 804 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 804 cttaaggatt taataccaga 20 <210> 805 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 805 tctcttaagg atttaatacc 20 <210> 806 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 806 gacagtgact ttaagataaa 20 <210> 807 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 807 tgtgattgta tgtttaatct 20 <210> 808 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 808 aggttatgtg attgtatgtt 20 <210> 809 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 809 ctttaaggtt atgtgattgt 20 <210> 810 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 810 ggtattcttt aaggttatgt 20 <210> 811 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 811 attgattccc acatcacaaa 20 <210> 812 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 812 ctaaaattga ttcccacatc 20 <210> 813 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 813 ccatctaaaa ttgattccca 20 <210> 814 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 814 ttgtgatatt agctcatatg 20 <210> 815 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 815 actagttttt taaactggga 20 <210> 816 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 816 gtcaagttta gagttttaac 20 <210> 817 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 817 tatttagtca agtttagagt 20 <210> 818 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 818 gatttttaca catactctgt 20 <210> 819 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 819 ctgcttcatt aggtttcata 20 <210> 820 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 820 caccagcctc ctaaaggaga 20 <210> 821 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 821 gacttcttaa ctctatatat 20 <210> 822 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 822 ctagacttct taactctata 20 <210> 823 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 823 gacctagact tcttaactct 20 <210> 824 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 824 ggaagcagac ctagacttct 20 <210> 825 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 825 tctggaagca gacctagact 20 <210> 826 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 826 tcttctggaa gcagacctag 20 <210> 827 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 827 ctaatcttta gggatttagg 20 <210> 828 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 828 tgtatctaat ctttagggat 20 <210> 829 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 829 taacttgggc actatatcct 20 <210> 830 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 830 attgacaaag gtaggtcacc 20 <210> 831 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 831 atatgacatg tatattggat 20 <210> 832 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 832 ttttgtactt ttctggaaca 20 <210> 833 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 833 tagtctgtgg tcctgaaaat 20 <210> 834 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 834 agcttagtct gtggtcctga 20 <210> 835 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 835 gacagcttag tctgtggtcc 20 <210> 836 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 836 gtattctggc cctaaaaaaa 20 <210> 837 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 837 attttggtat tctggcccta 20 <210> 838 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 838 gaaattgtcc aatttttggg 20 <210> 839 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 839 tttgcatttg aaattgtcca 20 <210> 840 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 840 ggaagcaact catatattaa 20 <210> 841 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 841 tatcagaaaa agatacctga 20 <210> 842 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 842 ataatagcta ataatgtggg 20 <210> 843 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 843 tgcagataat agctaataat 20 <210> 844 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 844 tgtcattgca gataatagct 20 <210> 845 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 845 taaaagttgt cattgcagat 20 <210> 846 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 846 cggattttta aaagttgtca 20 <210> 847 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 847 gggattcgga tttttaaaag 20 <210> 848 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 848 tttgggattc ggatttttaa 20 <210> 849 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 849 acgcttattt gggattcgga 20 <210> 850 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 850 tttaagagat ttacaagtca 20 <210> 851 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 851 gactacctgt ttttaaaagc 20 <210> 852 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 852 tatggtgact acctgttttt 20 <210> 853 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 853 actttgctgt attataaact 20 <210> 854 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 854 attgtattta actttgctgt 20 <210> 855 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 855 gagcaactaa cttaataggt 20 <210> 856 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 856 gaaatgagca actaacttaa 20 <210> 857 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 857 aatcaaagaa atgagcaact 20 <210> 858 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 858 accttcttcc acctgagtt 20 <210> 859 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 859 cacgaatgta accttcttcc 20 <210> 860 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 860 ttaacttgca cgaatgtaac 20 <210> 861 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 861 tatatatacc aatatttgcc 20 <210> 862 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 862 tcttaactct atatatacca 20 <210> 863 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 863 ctttaagtga agttacttct 20 <210> 864 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 864 tctacttact ttaagtgaag 20 <210> 865 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 865 gaaccctctt tattttctac 20 <210> 866 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 866 acataaacat gaaccctctt 20 <210> 867 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 867 ccacattgaa aacataaaca 20 <210> 868 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 868 gcatgcctta gaaatatttt 20 <210> 869 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 869 caatgcaaca aagtatttca 20 <210> 870 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 870 ctggagatta tttttcttgg 20 <210> 871 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 871 ttcatatata acattaggga 20 <210> 872 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 872 tcagtgtttt catataac 20 <210> 873 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 873 gacatagtgt tctagattgt 20 <210> 874 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 874 caatagtgta atgacatagt 20 <210> 875 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 875 ttacttacct tcagtaattt 20 <210> 876 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 876 atcttttcca tttactgtat 20 <210> 877 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 877 agaaaaagcc cagcatattt 20 <210> 878 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 878 gtatgcttct ttcaaatagc 20 <210> 879 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 879 ccttcccctt gtatgcttct 20 <210> 880 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 880 cctgtaacac tatcataatc 20 <210> 881 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 881 tgacttacct gattttctat 20 <210> 882 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 882 gatgggacat accattaaaa 20 <210> 883 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 883 gtgaaagatg ggacatacca 20 <210> 884 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 884 cctgtgtgaa agatgggaca 20 <210> 885 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 885 cattggctgc tatgaattaa 20 <210> 886 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 886 gatgacattg gctgctatga 20 <210> 887 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 887 gagaaacatg atctaatttg 20 <210> 888 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 888 atggaaagct attgtgtggt 20 <210> 889 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 889 gtctaaagag ccaatatgag 20 <210> 890 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 890 aatcttggtc taaagagcca 20 <210> 891 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 891 ggagcttgag atttcacttg 20 <210> 892 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 892 catcagattt agtaatagga 20 <210> 893 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 893 gttattacat cagattagt 20 <210> 894 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 894 cagcaggaat gcctagaatc 20 <210> 895 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 895 ctccttagac aggttttacc 20 <210> 896 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 896 gtctattctc cttagacagg 20 <210> 897 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 897 atgaatgatt gaatgtagtc 20 <210> 898 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 898 atatgaaggc tgagactgct 20 <210> 899 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 899 ataaattata tgaaggctga 20 <210> 900 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 900 atatttaaga acagacatgt 20 <210> 901 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 901 atttgtaaca gttactactt 20 <210> 902 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 902 cacagcttat ttgtaacagt 20 <210> 903 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 903 gtgactaatg ctaggagtgg 20 <210> 904 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 904 gaatagagtg actaatgcta 20 <210> 905 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 905 atgagagaat agagtgacta 20 <210> 906 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 906 tatactgtat gtctgagttt 20 <210> 907 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 907 aactaattca ttataagcca 20 <210> 908 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 908 tttggatttt aaacatctgt 20 <210> 909 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 909 tgtatgtgct ttttggattt 20 <210> 910 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 910 catggatttt tgtatgtgct 20 <210> 911 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 911 tcattcatgg atttttgtat 20 <210> 912 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 912 acttagacat cattcatgga 20 <210> 913 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 913 gtgagtactt agacatcatt 20 <210> 914 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 914 tttataagtg agtacttaga 20 <210> 915 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 915 gtcttctact ttataagtga 20 <210> 916 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 916 atgaatgtct tctactttat 20 <210> 917 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 917 caaatagtac tgagcattta 20 <210> 918 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 918 ttagaagatt tggagctaca 20 <210> 919 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 919 tcactattag aagatttgga 20 <210> 920 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 920 gggttacact cactattaga 20 <210> 921 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 921 acttacctgt cagcctttta 20 <210> 922 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 922 cttaccagaa ttaagtgagt 20 <210> 923 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 923 aatacaagta caaatgggtt 20 <210> 924 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 924 ctggtaaata caagtacaaa 20 <210> 925 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 925 ggattgctgg taaatacaag 20 <210> 926 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 926 tcattttaag gattgctggt 20 <210> 927 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 927 agttagtagg aagcttcatt 20 <210> 928 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 928 gctattgagt tagtaggaag 20 <210> 929 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 929 agcatggttc ttaataactt 20 <210> 930 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 930 ctttgtagaa aaagacagga 20 <210> 931 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 931 acctggcctt tggtatttgc 20 <210> 932 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 932 agtctttata tggataaact 20 <210> 933 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 933 cgtcattggt agaggaatat 20 <210> 934 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 934 gattatcctt tctataatgc 20 <210> 935 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 935 gtcttgaatc ccttgatcat 20 <210> 936 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 936 ggtgcaacta attgagttgt 20 <210> 937 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 937 gtgtttttta ttggtgcaac 20 <210> 938 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 938 attctcctga aaagaaaagt 20 <210> 939 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 939 atgccaccac caggctccta 20 <210> 940 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 940 atatccttta acaaatgggt 20 <210> 941 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 941 gcactatatc ctttaacaaa 20 <210> 942 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 942 acttgggcac tatatccttt 20 <210> 943 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 943 gaaacatgtc ctatgagagt 20 <210> 944 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 944 ttgagcactt taagcaaagt 20 <210> 945 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 945 ggaatttgag cactttaagc 20 <210> 946 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 946 tagattagac aactgtgagt 20 <210> 947 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 947 aaaatgaagg tcaagtttga 20 <210> 948 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 948 gtgaaagcaa aatgaaggtc 20 <210> 949 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 949 gtattgtgaa agcaaaatga 20 <210> 950 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 950 tggagagtat agtattgtga 20 <210> 951 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 951 gagatttaca agtcaaaaat 20 <210> 952 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 952 atttaacttt gctgtattat 20 <210> 953 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 953 atcaatgcta aatgaaatca 20 <210> 954 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 954 tattttctgg agattatttt 20 <210> 955 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 955 aaaatgaata ttggcaattc 20 <210> 956 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 956 aatgcctaga atcaataaaa 20 <210> 957 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 957 gtaaatattt gtagattagc 20 <210> 958 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 958 acaaatgtgt aattgtttga 20 <210> 959 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 959 tactaacaaa tgtgtaattg 20 <210> 960 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 960 tgataagtat atttaagaac 20 <210> 961 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 961 ttaacttcca attaattgat 20 <210> 962 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 962 tctgttattt tatcttgctt 20 <210> 963 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 963 atcacaatcc tttttattaa 20 <210> 964 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 964 agagacttga gtaataataa 20 <210> 965 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 965 aacaaaatga aacatgtcct 20 <210> 966 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 966 gattttcaat ttcaagcaac 20 <210> 967 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 967 agcttaattg tgaacatttt 20 <210> 968 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 968 gaaggagctt aattgtgaac 20 <210> 969 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 969 caataaaaag aaggagctta 20 <210> 970 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 970 gaacaataaa aagaaggagc 20 <210> 971 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 971 ctggaggaaa taactagagg 20 <210> 972 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 972 attctggagg aaataactag 20 <210> 973 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 973 tcaaatgatg aattgtcttg 20 <210> 974 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 974 ttgattttgg ctctggagat 20 <210> 975 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 975 gcaaatcttg attttggctc 20 <210> 976 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 976 atagcaaatc ttgattttgg 20 <210> 977 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 977 cgtctaacat agcaaatctt 20 <210> 978 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 978 tggctaaaat ttttacatcg 20 <210> 979 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 979 ccattggcta aaatttttac 20 <210> 980 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 980 aggaggccat tggctaaaat 20 <210> 981 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 981 tgaaggaggc cattggctaa 20 <210> 982 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 982 gtcccaactg aaggaggcca 20 <210> 983 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 983 ccatgtccca actgaaggag 20 <210> 984 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 984 agaccatgtc ccaactgaag 20 <210> 985 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 985 tttaagacca tgtcccaact 20 <210> 986 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 986 gtctttaaga ccatgtccca 20 <210> 987 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 987 ggacaaagtc tttaagacca 20 <210> 988 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 988 tcttatggac aaagtcttta 20 <210> 989 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 989 tatgtcatta atttggccct 20 <210> 990 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 990 gatcaaatat gttgagtttt 20 <210> 991 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 991 gactgatcaa atatgttgag 20 <210> 992 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 992 tcttctttga tttcactggt 20 <210> 993 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 993 cttctcagtt ccttttcttc 20 <210> 994 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 994 gttcttctca gttccttttc 20 <210> 995 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 995 tgtagttctt ctcagttcct 20 <210> 996 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 996 tatatgtagt tcttctcagt 20 <210> 997 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 997 gtttatatgt agttcttctc 20 <210> 998 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 998 tgtagtttat atgtagttct 20 <210> 999 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 999 gacttgtagt ttatatgtag 20 <210> 1000 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1000 tcatttttga cttgtagttt 20 <210> 1001 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1001 cctcttcatt tttgacttgt 20 <210> 1002 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1002 tctttacctc ttcatttttg 20 <210> 1003 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1003 catattcttt acctcttcat 20 <210> 1004 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1004 gtgacatatt ctttacctct 20 <210> 1005 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1005 gagttcaagt gacatattct 20 <210> 1006 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1006 tgagttgagt tcaagtgaca 20 <210> 1007 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1007 agttttgagt tgagttcaag 20 <210> 1008 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1008 tcaagttttg agttgagttc 20 <210> 1009 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1009 ggaggctttc aagttttgag 20 <210> 1010 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1010 tttcttctag gaggctttca 20 <210> 1011 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1011 attttttctt ctaggaggct 20 <210> 1012 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1012 gtagaatttt ttcttctagg 20 <210> 1013 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1013 gctcttctaa atatttcact 20 <210> 1014 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1014 agttagttag ttgctcttct 20 <210> 1015 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1015 caggttgatt ttgaattaag 20 <210> 1016 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1016 tttcaggttg attttgaatt 20 <210> 1017 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1017 ggagtttcag gttgattttg 20 <210> 1018 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1018 gttctggagt ttcaggttga 20 <210> 1019 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1019 ttaagtgaag ttacttctgg 20 <210> 1020 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1020 tgctattatc ttgtttttct 20 <210> 1021 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1021 ggtctttgat gctattatct 20 <210> 1022 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1022 gaaggtcttt gatgctatta 20 <210> 1023 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1023 ctggagaagg tctttgatgc 20 <210> 1024 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1024 ctgagctgat tttctatttc 20 <210> 1025 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1025 atttctgtgg gttcttgaat 20 <210> 1026 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1026 gagaaatttc tgtgggttct 20 <210> 1027 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1027 gatagagaaa tttctgtggg 20 <210> 1028 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1028 cttggaagat agagaaattt 20 <210> 1029 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1029 gtgctcttgg cttggaagat 20 <210> 1030 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1030 cttggtgctc ttggcttgga 20 <210> 1031 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1031 gggagtagtt cttggtgctc 20 <210> 1032 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1032 ctgaagaaag ggagtagttc 20 <210> 1033 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1033 atcatgtttt acatttctta 20 <210> 1034 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1034 gccatcatgt tttacatttc 20 <210> 1035 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1035 gaatgccatc atgttttaca 20 <210> 1036 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1036 caggaatgcc atcatgtttt 20 <210> 1037 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1037 aaagtcaatg tgacttagta 20 <210> 1038 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1038 aaggtatagt gatacctcat 20 <210> 1039 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1039 actgttttct tctggaagca 20 <210> 1040 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1040 aaataaggta tagtgatacc 20 <210> 1041 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1041 acaaataagg tatagtgata 20 <210> 1042 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1042 taacaaataa ggtatagtga 20 <210> 1043 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1043 tttaacaaat aaggtatagt 20 <210> 1044 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1044 atatatttta acaaataagg 20 <210> 1045 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1045 cagtatatat tttaacaaat 20 <210> 1046 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1046 tacagtatat attttaacaa 20 <210> 1047 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1047 tatacagtat atattttaac 20 <210> 1048 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1048 atagtattaa gtgttaaaat 20 <210> 1049 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1049 tcatagtatt aagtgttaaa 20 <210> 1050 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1050 gttttcatag tattaagtgt 20 <210> 1051 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1051 attatttgtt ttcatagtat 20 <210> 1052 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1052 ctttacaatt atttgttttc 20 <210> 1053 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1053 attcctttac aattatttgt 20 <210> 1054 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1054 agattccttt acaattattt 20 <210> 1055 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1055 caagatcct ttacaattat 20 <210> 1056 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1056 gacaagatc ctttacaatt 20 <210> 1057 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1057 ctgacaagat tcctttacaa 20 <210> 1058 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1058 aatctgacaa gattccttta 20 <210> 1059 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1059 gtaatctgac aagattcctt 20 <210> 1060 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1060 ctgtaatctg acaagattcc 20 <210> 1061 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1061 tactgtaatc tgacaagatt 20 <210> 1062 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1062 cttactgtaa tctgacaaga 20 <210> 1063 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1063 ttcttactgt aatctgacaa 20 <210> 1064 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1064 cattcttact gtaatctgac 20 <210> 1065 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1065 ttcattctta ctgtaatctg 20 <210> 1066 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1066 tgttcattct tactgtaatc 20 <210> 1067 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1067 tatgttcatt cttactgtaa 20 <210> 1068 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1068 aatatgttca ttcttactgt 20 <210> 1069 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1069 acaaatatgt tcattcttac 20 <210> 1070 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1070 cgatgccaca aatatgttca 20 <210> 1071 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1071 ctcgatgcca caaatatgtt 20 <210> 1072 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1072 aactcgatgc cacaaatatg 20 <210> 1073 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1073 ttaactcgat gccacaaata 20 <210> 1074 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1074 ctttaactcg atgccacaaa 20 <210> 1075 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1075 aactttaact cgatgccaca 20 <210> 1076 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1076 aatataaact ttaactcgat 20 <210> 1077 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1077 gaaatataaa ctttaactcg 20 <210> 1078 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1078 gggaaatata aactttaact 20 <210> 1079 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1079 gaatcacagc atatttaggg 20 <210> 1080 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1080 tagaatcaca gcatatttag 20 <210> 1081 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1081 gtattagaat cacagcatat 20 <210> 1082 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1082 atgtattaga atcacagcat 20 <210> 1083 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1083 gaatgtatta gaatcacagc 20 <210> 1084 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1084 acgaatgtat tagaatcaca 20 <210> 1085 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1085 acacgaatgt attagaatca 20 <210> 1086 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1086 ctacacgaat gtattagaat 20 <210> 1087 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1087 acctacacga atgtattaga 20 <210> 1088 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1088 aaacctacac gaatgtatta 20 <210> 1089 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1089 gaaaacctac acgaatgtat 20 <210> 1090 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1090 ttgaaaacct acacgaatgt 20 <210> 1091 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1091 acttgaaaac ctacacgaat 20 <210> 1092 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1092 ctacttgaaa acctacacga 20 <210> 1093 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1093 tatttctact tgaaaaccta 20 <210> 1094 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1094 tttatttcta cttgaaaacc 20 <210> 1095 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1095 ggtttatttc tacttgaaaa 20 <210> 1096 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1096 gaggtttatt tctacttgaa 20 <210> 1097 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1097 ttacgaggtt tatttctact 20 <210> 1098 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1098 tgttacgagg tttatttcta 20 <210> 1099 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1099 cttgttacga ggtttatttc 20 <210> 1100 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1100 aacttgttac gaggtttatt 20 <210> 1101 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1101 gtaacttgtt acgaggttta 20 <210> 1102 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1102 cagtaacttg ttacgaggtt 20 <210> 1103 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1103 ttcagtaact tgttacgagg 20 <210> 1104 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1104 cgttcagtaa cttgttacga 20 <210> 1105 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1105 cttgtcaggc tgtttaaacg 20 <210> 1106 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1106 tgcttgtcag gctgtttaaa 20 <210> 1107 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1107 tatacatgct tgtcaggctg 20 <210> 1108 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1108 tatatacatg cttgtcaggc 20 <210> 1109 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1109 catatataca tgcttgtcag 20 <210> 1110 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1110 tggaactgtt ttcttctgga 20 <210> 1111 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1111 cgtggaactg ttttcttctg 20 <210> 1112 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1112 tcttgatttt caatttcaag 20 <210> 1113 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1113 ttttatcttg attttcaatt 20 <210> 1114 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1114 catttttatc ttgattttca 20 <210> 1115 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1115 attgtgaaca tttttatctt 20 <210> 1116 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1116 taattgtgaa catttttatc 20 <210> 1117 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1117 aagaaggagc ttaattgtga 20 <210> 1118 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1118 aaaagaagga gcttaattgt 20 <210> 1119 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1119 taaaaagaag gagcttaatt 20 <210> 1120 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1120 taactagagg aacaataaaa 20 <210> 1121 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1121 aataactaga ggaacaataa 20 <210> 1122 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1122 gaaataacta gaggaacaat 20 <210> 1123 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1123 aggaaataac tagaggaaca 20 <210> 1124 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1124 ggaggaaata actagaggaa 20 <210> 1125 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1125 caattctgga ggaaataact 20 <210> 1126 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1126 atcaattctg gaggaaataa 20 <210> 1127 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1127 cttgatcaat tctggaggaa 20 <210> 1128 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1128 gtcttgatca attctggagg 20 <210> 1129 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1129 ttgtcttgat caattctgga 20 <210> 1130 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1130 aattgtcttg atcaattctg 20 <210> 1131 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1131 tgaattgtct tgatcaattc 20 <210> 1132 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1132 gatgaattgt cttgatcaat 20 <210> 1133 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1133 atgatgaatt gtcttgatca 20 <210> 1134 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1134 aaatgatgaa ttgtcttgat 20 <210> 1135 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1135 aatcaaatga tgaattgtct 20 <210> 1136 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1136 agaatcaaat gatgaattgt 20 <210> 1137 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1137 tagagaatca aatgatgaat 20 <210> 1138 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1138 ctggagatag agaatcaaat 20 <210> 1139 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1139 ctctggagat agagaatcaa 20 <210> 1140 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1140 ttggctctgg agatagagaa 20 <210> 1141 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1141 attttggctc tggagataga 20 <210> 1142 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1142 aatcttgatt ttggctctgg 20 <210> 1143 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1143 acatagcaaa tcttgatttt 20 <210> 1144 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1144 taacatagca aatcttgatt 20 <210> 1145 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1145 tctaacatag caaatcttga 20 <210> 1146 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1146 attggctaaa atttttacat 20 <210> 1147 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1147 ggccattggc taaaattttt 20 <210> 1148 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1148 gaggccattg gctaaaattt 20 <210> 1149 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1149 actgaaggag gccattggct 20 <210> 1150 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1150 caactgaagg aggccattgg 20 <210> 1151 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1151 cccaactgaa ggaggccatt 20 <210> 1152 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1152 atgtcccaac tgaaggaggc 20 <210> 1153 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1153 taagaccatg tcccaactga 20 <210> 1154 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1154 aagtctttaa gaccatgtcc 20 <210> 1155 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1155 caaagtcttt aagaccatgt 20 <210> 1156 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1156 tatggacaaa gtctttaaga 20 <210> 1157 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1157 cgtcttatgg acaaagtctt 20 <210> 1158 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1158 aatatgtcat taatttggcc 20 <210> 1159 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1159 gaaatatgtc attaatttgg 20 <210> 1160 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1160 tttgaaatat gtcattaatt 20 <210> 1161 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1161 agtttttgaa atatgtcatt 20 <210> 1162 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1162 tgagtttttg aaatatgtca 20 <210> 1163 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1163 caaatatgtt gagtttttga 20 <210> 1164 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1164 ctgatcaaat atgttgagtt 20 <210> 1165 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1165 aagactgatc aaatatgttg 20 <210> 1166 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1166 taaaaagact gatcaaatat 20 <210> 1167 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1167 cataaaaaga ctgatcaaat 20 <210> 1168 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1168 atcataaaaa gactgatcaa 20 <210> 1169 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1169 tagatcataa aaagactgat 20 <210> 1170 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1170 gatagatcat aaaaagactg 20 <210> 1171 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1171 gcgatagatc ataaaaagac 20 <210> 1172 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1172 cagcgataga tcataaaaag 20 <210> 1173 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1173 tgcagcgata gatcataaaa 20 <210> 1174 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1174 tttgcagcga tagatcataa 20 <210> 1175 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1175 ggtttgcagc gatagatcat 20 <210> 1176 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1176 ctggtttgca gcgatagatc 20 <210> 1177 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1177 cactggtttg cagcgataga 20 <210> 1178 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1178 atttcactgg tttgcagcga 20 <210> 1179 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1179 ttgatttcac tggtttgcag 20 <210> 1180 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1180 cttcttcttt gatttcactg 20 <210> 1181 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1181 gttccttttc ttcttctttg 20 <210> 1182 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1182 ttgtatgttc acctctgtta 20 <210> 1183 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1183 cttgtatgtt cacctctgtt 20 <210> 1184 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1184 gccacttgta tgttcacctc 20 <210> 1185 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1185 tgccacttgt atgttcacct 20 <210> 1186 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1186 atgccacttg tatgttcacc 20 <210> 1187 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1187 catgccactt gtatgttcac 20 <210> 1188 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1188 acagccact tgtatgttca 20 <210> 1189 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1189 tacatgccac ttgtatgttc 20 <210> 1190 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1190 gcatacatgc cacttgtatg 20 <210> 1191 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1191 ggcatacatg ccacttgtat 20 <210> 1192 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1192 tggcatacat gccacttgta 20 <210> 1193 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1193 atggcataca tgccacttgt 20 <210> 1194 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1194 tctgatggca tacatgccac 20 <210> 1195 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1195 gtctgatggc atacatgcca 20 <210> 1196 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1196 gggtctgatg gcatacatgc 20 <210> 1197 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1197 tgggtctgat ggcatacatg 20 <210> 1198 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1198 ctgggtctga tggcatacat 20 <210> 1199 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1199 gagagttgct gggtctgatg 20 <210> 1200 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1200 tgagagttgc tgggtctgat 20 <210> 1201 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1201 ttgagagttg ctgggtctga 20 <210> 1202 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1202 cttgagagtt gctgggtctg 20 <210> 1203 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1203 acttgagagt tgctgggtct 20 <210> 1204 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1204 aacttgagag ttgctgggtc 20 <210> 1205 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1205 gaaaaacttg agagttgctg 20 <210> 1206 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1206 tgaaaaactt gagagttgct 20 <210> 1207 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1207 atgaaaaact tgagagttgc 20 <210> 1208 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1208 catgaaaaac ttgagagttg 20 <210> 1209 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1209 gtagacatga aaaacttgag 20 <210> 1210 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1210 cagtagacat gaaaaacttg 20 <210> 1211 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1211 taacatcaca gtagacatga 20 <210> 1212 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1212 ataacatcac agtagacatg 20 <210> 1213 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1213 tataacatca cagtagacat 20 <210> 1214 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1214 atataacatc acagtagaca 20 <210> 1215 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1215 gatataacat cacagtagac 20 <210> 1216 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1216 tgatataaca tcacagtaga 20 <210> 1217 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1217 ctgatataac atcacagtag 20 <210> 1218 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1218 cctgatataa catcacagta 20 <210> 1219 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1219 acctgatata acatcacagt 20 <210> 1220 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1220 tacctgatat aacatcacag 20 <210> 1221 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1221 ctacctgata taacatcaca 20 <210> 1222 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1222 actacctgat ataacatcac 20 <210> 1223 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1223 gactacctga tataacatca 20 <210> 1224 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1224 ggactacctg atataacatc 20 <210> 1225 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1225 tggactacct gatataacat 20 <210> 1226 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1226 atggactacc tgatataaca 20 <210> 1227 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1227 catggactac ctgatataac 20 <210> 1228 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1228 ccatggacta cctgatataa 20 <210> 1229 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1229 tccatggact acctgatata 20 <210> 1230 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1230 gtccatggac tacctgatat 20 <210> 1231 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1231 tgtccatgga ctacctgata 20 <210> 1232 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1232 atgtccatgg actacctgat 20 <210> 1233 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1233 aatgtccatg gactacctga 20 <210> 1234 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1234 taatgtccat ggactacctg 20 <210> 1235 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1235 ttaatgtcca tggactacct 20 <210> 1236 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1236 attaatgtcc atggactacc 20 <210> 1237 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1237 aattaatgtc catggactac 20 <210> 1238 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1238 gttgaattaa tgtccatgga 20 <210> 1239 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1239 tgttgaatta atgtccatgg 20 <210> 1240 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1240 atgttgaatt aatgtccat 20 <210> 1241 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1241 gatgttgaat taatgtccat 20 <210> 1242 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1242 cgatgttgaa ttaatgtcca 20 <210> 1243 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1243 tcgatgttga attaatgtcc 20 <210> 1244 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1244 ttcgatgttg aattaatgtc 20 <210> 1245 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1245 attcgatgtt gaattaatgt 20 <210> 1246 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1246 tattcgatgt tgaattaatg 20 <210> 1247 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1247 ctattcgatg ttgaattaat 20 <210> 1248 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1248 atctattcga tgttgaatta 20 <210> 1249 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1249 catctattcg atgttgaatt 20 <210> 1250 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1250 ccatctattc gatgttgaat 20 <210> 1251 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1251 tccatctatt cgatgttgaa 20 <210> 1252 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1252 atccatctat tcgatgttga 20 <210> 1253 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1253 gatccatcta ttcgatgttg 20 <210> 1254 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1254 tgatccatct attcgatgtt 20 <210> 1255 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1255 gtgatccatc tattcgatgt 20 <210> 1256 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1256 tgtgatccat ctattcgatg 20 <210> 1257 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1257 ttgtgatcca tctattcgat 20 <210> 1258 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1258 tttaggttgt tttctccaca 20 <210> 1259 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1259 atttaggttg ttttctccac 20 <210> 1260 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1260 taccatttag gttgttttct 20 <210> 1261 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1261 gttattatta ccatttaggt 20 <210> 1262 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1262 tgtttatattt accatttagg 20 <210> 1263 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1263 ttgttatatt taccatttag 20 <210> 1264 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1264 tttgttatat ttaccattta 20 <210> 1265 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1265 tggtttgtta tatttaccat 20 <210> 1266 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1266 ctcttggttt gttatattta 20 <210> 1267 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1267 gctcttggtt tgttatattt 20 <210> 1268 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1268 tgctcttggt ttgttatatt 20 <210> 1269 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1269 ttttgctctt ggtttgttat 20 <210> 1270 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1270 attttgctct tggtttgtta 20 <210> 1271 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1271 gattttgctc ttggtttgtt 20 <210> 1272 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1272 agattttgct cttggtttgt 20 <210> 1273 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1273 tagattttgc tcttggtttg 20 <210> 1274 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1274 ctctctggct tagattttgc 20 <210> 1275 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1275 cctctctggc ttagattttg 20 <210> 1276 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1276 tcctctctgg cttagatttt 20 <210> 1277 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1277 cttctcctct ctggcttaga 20 <210> 1278 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1278 tcttctcctc tctggcttag 20 <210> 1279 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1279 ctcttctcct ctctggctta 20 <210> 1280 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1280 taatcctctt ctcctctctg 20 <210> 1281 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1281 ataatcctct tctcctctct 20 <210> 1282 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1282 gataatcctc ttctcctctc 20 <210> 1283 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1283 agataatcct cttctcctct 20 <210> 1284 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1284 aagataatcc tcttctcctc 20 <210> 1285 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1285 caagataatc ctcttctcct 20 <210> 1286 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1286 ccaagataat cctcttctcc 20 <210> 1287 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1287 tccaagataa tcctcttctc 20 <210> 1288 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1288 ttccaagata atcctcttct 20 <210> 1289 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1289 cttccaagat aatcctcttc 20 <210> 1290 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1290 acttccaaga taatcctctt 20 <210> 1291 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1291 gacttccaag ataatcctct 20 <210> 1292 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1292 agacttccaa gataatcctc 20 <210> 1293 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1293 gagacttcca agataatcct 20 <210> 1294 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1294 tgagacttcc aagataatcc 20 <210> 1295 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1295 ttgagacttc caagataatc 20 <210> 1296 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1296 tttgagactt ccaagataat 20 <210> 1297 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1297 ttttgagact tccaagataa 20 <210> 1298 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1298 attttgagac ttccaagata 20 <210> 1299 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1299 ccattttgag acttccaaga 20 <210> 1300 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1300 cttccatttt gagacttcca 20 <210> 1301 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1301 taaccttcca ttttgagact 20 <210> 1302 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1302 ataaccttcc attttgagac 20 <210> 1303 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1303 tataaccttc cattttgaga 20 <210> 1304 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1304 gtataacctt ccattttgag 20 <210> 1305 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1305 gagtataacc ttccattttg 20 <210> 1306 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1306 agagtataac cttccatttt 20 <210> 1307 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1307 ttatagagta taaccttcca 20 <210> 1308 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1308 gattttatag agtataacct 20 <210> 1309 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1309 tgattttata gagtataacc 20 <210> 1310 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1310 ttgattttat agagtataac 20 <210> 1311 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1311 gttgatttta tagagtataa 20 <210> 1312 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1312 tttggttgat tttatagagt 20 <210> 1313 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1313 ggatcaacat tttggttgat 20 <210> 1314 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1314 tggatcaaca ttttggttga 20 <210> 1315 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1315 atggatcaac attttggttg 20 <210> 1316 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1316 aatctgttgg atggatcaac 20 <210> 1317 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1317 gaatctgttg gatggatcaa 20 <210> 1318 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1318 ttctgaatct gttggatgga 20 <210> 1319 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1319 tttctgaatc tgttggatgg 20 <210> 1320 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1320 ctttctgaat ctgttggatg 20 <210> 1321 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1321 aaagctttct gaatctgttg 20 <210> 1322 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1322 ttgcctcagt tcattcaaag 20 <210> 1323 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1323 aaatttgcct cagttcattc 20 <210> 1324 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1324 cttttaaatt tgcctcagtt 20 <210> 1325 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1325 tattgccttt taaatttgcc 20 <210> 1326 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1326 ttattgcctt ttaaatttgc 20 <210> 1327 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1327 ttaaattatt gccttttaaa 20 <210> 1328 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1328 tttaaattat tgccttttaa 20 <210> 1329 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1329 gtttaaatta ttgcctttta 20 <210> 1330 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1330 tgtttaaatt attgcctttt 20 <210> 1331 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1331 atgtttaaat tattgccttt 20 <210> 1332 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1332 tttaataagt tcacctattg 20 <210> 1333 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1333 atttaataag ttcacctatt 20 <210> 1334 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1334 tattataataa gttcacctat 20 <210> 1335 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1335 ttatttaata agttcaccta 20 <210> 1336 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1336 gttatttaat aagttcacct 20 <210> 1337 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1337 agttatttaa taagttcacc 20 <210> 1338 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1338 aaaagttatt taataagttc 20 <210> 1339 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1339 tatttagaaa agttatttaa 20 <210> 1340 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1340 taaaagtctc taaatttttt 20 <210> 1341 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1341 ataaaagtct ctaaattttt 20 <210> 1342 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1342 aataaaagtc tctaaatttt 20 <210> 1343 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1343 ttagctcata tgatgccttt 20 <210> 1344 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1344 attagctcat atgatgcctt 20 <210> 1345 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1345 atattagctc atatgatgcc 20 <210> 1346 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1346 gatattagct catatgatgc 20 <210> 1347 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1347 tgatattagc tcatatgatg 20 <210> 1348 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1348 gtgatattag ctcatatgat 20 <210> 1349 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1349 gaaagttgtg atattagctc 20 <210> 1350 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1350 ggaaagttgt gatattagct 20 <210> 1351 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1351 gggaaagttg tgatattagc 20 <210> 1352 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1352 tgggaaagtt gtgatattag 20 <210> 1353 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1353 ctgggaaagt tgtgatatta 20 <210> 1354 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1354 actgggaaag ttgtgatatt 20 <210> 1355 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1355 aactgggaaa gttgtgatat 20 <210> 1356 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1356 aaactgggaa agttgtgata 20 <210> 1357 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1357 taaactggga aagttgtgat 20 <210> 1358 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1358 ttaaactggg aaagttgtga 20 <210> 1359 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1359 tttaaactgg gaaagttgtg 20 <210> 1360 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1360 ttttaaactg ggaaagttgt 20 <210> 1361 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1361 gttttttaaa ctgggaaagt 20 <210> 1362 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1362 agttttttaa actgggaaag 20 <210> 1363 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1363 tagtttttta aactgggaaa 20 <210> 1364 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1364 gtactagttt tttaaactgg 20 <210> 1365 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1365 agagtactag ttttttaaac 20 <210> 1366 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1366 caagagtact agttttttaa 20 <210> 1367 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1367 tttaacaaga gtactagttt 20 <210> 1368 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1368 ttttaacaag agtactagtt 20 <210> 1369 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1369 gttttaacaa gagtactagt 20 <210> 1370 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1370 agttttaaca agagtactag 20 <210> 1371 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1371 gagttttaac aagagtacta 20 <210> 1372 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1372 agagttttaa caagagtact 20 <210> 1373 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1373 tttagagttt taacaagagt 20 <210> 1374 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1374 gtttagagtt ttaacaagag 20 <210> 1375 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1375 aagtttagag ttttaacaag 20 <210> 1376 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1376 caagtttaga gttttaacaa 20 <210> 1377 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1377 tagtcaagtt tagagtttta 20 <210> 1378 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1378 ttagtcaagt ttagagtttt 20 <210> 1379 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1379 tttagtcaag tttagagttt 20 <210> 1380 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1380 tgtatttagt caagtttaga 20 <210> 1381 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1381 ctgtatttag tcaagtttag 20 <210> 1382 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1382 tctgtatttta gtcaagttta 20 <210> 1383 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1383 gtcctctgta tttagtcaag 20 <210> 1384 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1384 agtcctctgt atttagtcaa 20 <210> 1385 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1385 cagtcctctg tatttagtca 20 <210> 1386 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1386 ccagtcctct gtatttagtc 20 <210> 1387 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1387 accagtcctc tgtatttagt 20 <210> 1388 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1388 taccagtcct ctgtatttag 20 <210> 1389 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1389 ttaccagtcc tctgtatttta 20 <210> 1390 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1390 attaccagtc ctctgtattt 20 <210> 1391 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1391 aattaccagt cctctgtatt 20 <210> 1392 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1392 caattaccag tcctctgtat 20 <210> 1393 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1393 acaattacca gtcctctgta 20 <210> 1394 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1394 tacaattacc agtcctctgt 20 <210> 1395 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1395 gtacaattac cagtcctctg 20 <210> 1396 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1396 tgtacaatta ccagtcctct 20 <210> 1397 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1397 ctgtacaatt accagtcctc 20 <210> 1398 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1398 actgtacaat taccagtcct 20 <210> 1399 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1399 aactgtacaa ttaccagtcc 20 <210> 1400 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1400 gaactgtaca attaccagtc 20 <210> 1401 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1401 agaactgtac aattaccagt 20 <210> 1402 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1402 taagaactgt acaattacca 20 <210> 1403 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1403 ttaagaactg tacaattacc 20 <210> 1404 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1404 tttaagaact gtacaattac 20 <210> 1405 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1405 acttgaaatt ataataggaa 20 <210> 1406 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1406 aaaaaactaa cttgaaatta 20 <210> 1407 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1407 gaaacaaaaa actaacttga 20 <210> 1408 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1408 gtgttttcat atataacatt 20 <210> 1409 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1409 aattttcagt gttttcatat 20 <210> 1410 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1410 aaaatgcaaa ttttcagtgt 20 <210> 1411 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1411 gtaattttca tataaaatgc 20 <210> 1412 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1412 gatttgtaat tttcatataa 20 <210> 1413 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1413 taaccgattt gtaattttca 20 <210> 1414 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1414 taatttaacc gatttgtaat 20 <210> 1415 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1415 ttgtataatt taaccgattt 20 <210> 1416 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1416 ctagattgta taatttaacc 20 <210> 1417 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1417 gtgttctaga ttgtataatt 20 <210> 1418 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1418 aatgacatag tgttctagat 20 <210> 1419 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1419 agtgtaatga catagtgttc 20 <210> 1420 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1420 ttacaatagt gtaatgacat 20 <210> 1421 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1421 ttcagtaatt tacaatagtg 20 <210> 1422 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1422 ttaccttcag taatttacaa 20 <210> 1423 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1423 ttaacttttt acttaccttc 20 <210> 1424 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1424 gaatagtttt aaattttttt 20 <210> 1425 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1425 acactggaga atagttttaa 20 <210> 1426 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1426 tttaaacact ggagaatagt 20 <210> 1427 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1427 tctgttttaa acactggaga 20 <210> 1428 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1428 gtattatttta atctgtttta 20 <210> 1429 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1429 ttactgtatt atttaatctg 20 <210> 1430 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1430 taaatctttt ccatttactg 20 <210> 1431 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1431 atgaataaat cttttccatt 20 <210> 1432 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1432 gcatattttc atatgaataa 20 <210> 1433 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1433 gcccagcata ttttcatatg 20 <210> 1434 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1434 aaaagaaaaa gcccagcata 20 <210> 1435 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1435 ctgaacttca attaaaagaa 20 <210> 1436 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1436 gattttctga acttcaatta 20 <210> 1437 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1437 tctaaaattt gattttctga 20 <210> 1438 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1438 actatctcta aaatttgatt 20 <210> 1439 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1439 ttaaattgta ctatctctaa 20 <210> 1440 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1440 acattttatt taaattgtac 20 <210> 1441 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1441 gtccttaaca ttttatttaa 20 <210> 1442 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1442 catatttttg tccttaacat 20 <210> 1443 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1443 tagcacatat ttttgtcctt 20 <210> 1444 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1444 tcaaatagca catatttttg 20 <210> 1445 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1445 cttctttcaa atagcacata 20 <210> 1446 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1446 cttgtatgct tctttcaaat 20 <210> 1447 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1447 attccttccc cttgtatgct 20 <210> 1448 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1448 ttggcaattc cttccccttg 20 <210> 1449 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1449 gaatattggc aattccttcc 20 <210> 1450 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1450 tgaaaaatga atattggcaa 20 <210> 1451 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1451 taatggattt gaaaaatgaa 20 <210> 1452 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1452 actaataatg gatttgaaaa 20 <210> 1453 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1453 cataatctaa atttttaaac 20 <210> 1454 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1454 cactatcata atctaaattt 20 <210> 1455 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1455 aatttcctgt aacactatca 20 <210> 1456 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1456 ctattaattt cctgtaacac 20 <210> 1457 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1457 cttttctatt aatttcctgt 20 <210> 1458 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1458 ctctttcttt tctattaatt 20 <210> 1459 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1459 agttgctttc ctctttcttt 20 <210> 1460 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1460 ttataagttg ctttcctctt 20 <210> 1461 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1461 gttggttata agttgctttc 20 <210> 1462 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1462 agtaggttgg ttataagttg 20 <210> 1463 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1463 tagagagtag gttggttata 20 <210> 1464 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1464 ggatatagag agtaggttgg 20 <210> 1465 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1465 agtctggata tagagagtag 20 <210> 1466 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1466 tacaaaagtc tggatataga 20 <210> 1467 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1467 gtttttctac aaaagtctgg 20 <210> 1468 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1468 ttacctgatt ttctatttct 20 <210> 1469 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1469 atactgactt acctgatttt 20 <210> 1470 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1470 ttaaaatact gacttacctg 20 <210> 1471 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1471 taccattaaa atactgactt 20 <210> 1472 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1472 ggacatacca ttaaaatact 20 <210> 1473 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1473 aaagatggga cataccatta 20 <210> 1474 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1474 agacctgtgt gaaagatggg 20 <210> 1475 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1475 tttacagacc tgtgtgaaag 20 <210> 1476 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1476 gtgtttttac agacctgtgt 20 <210> 1477 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1477 attcagtgtt tttacagacc 20 <210> 1478 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1478 ttaggattca gtgtttttac 20 <210> 1479 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1479 ataattttag gattcagtgt 20 <210> 1480 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1480 gcttgtaaat aattttagga 20 <210> 1481 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1481 gttaaagctt gtaaataatt 20 <210> 1482 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1482 tgttttatat ctcttgaaaa 20 <210> 1483 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1483 ttggtaataa tatttgtttt 20 <210> 1484 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1484 ggaaattggt aataatattt 20 <210> 1485 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1485 ttagtggaaa ttggtaataa 20 <210> 1486 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1486 tttgtttagt ggaaattggt 20 <210> 1487 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1487 ttatgtttgt ttagtggaaa 20 <210> 1488 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1488 taacattatg tttgtttagt 20 <210> 1489 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1489 actactaaca ttatgtttgt 20 <210> 1490 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1490 gcagcactac taacattatg 20 <210> 1491 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1491 ttttagcagc actactaaca 20 <210> 1492 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1492 aaacctttta gcagcactac 20 <210> 1493 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1493 gataaaaaac cttttagcag 20 <210> 1494 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1494 tagttgataa aaaacctttt 20 <210> 1495 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1495 caaaagtagt tgataaaaaa 20 <210> 1496 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1496 atggaaacca aaagtagttg 20 <210> 1497 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1497 aaagtatgga aaccaaaagt 20 <210> 1498 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1498 gaaggaaagt atggaaacca 20 <210> 1499 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1499 cataagaagg aaagtatgga 20 <210> 1500 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1500 taacatcata agaaggaaag 20 <210> 1501 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1501 gaataataac atcataagaa 20 <210> 1502 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1502 gaatttagaa taataacatc 20 <210> 1503 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1503 tataattgaa aagaatttag 20 <210> 1504 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1504 tagtaaaaga tataattgaa 20 <210> 1505 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1505 aatcatagta aaagatataa 20 <210> 1506 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1506 caggttcatt taatcatagt 20 <210> 1507 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1507 ctatagtaac attttgcttt 20 <210> 1508 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1508 gtatattact atagtaacat 20 <210> 1509 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1509 acaatgtata ttactatagt 20 <210> 1510 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1510 tagacacaat gtatattact 20 <210> 1511 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1511 tatttttaga cacaatgtat 20 <210> 1512 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1512 acacattttt atttttagac 20 <210> 1513 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1513 ttggtttctt cacacatttt 20 <210> 1514 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1514 ttcattgttt tggtttcttc 20 <210> 1515 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1515 cagaaattca ttgttttggt 20 <210> 1516 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1516 cttcttccaa ctcagaaatt 20 <210> 1517 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1517 tgatctaact cttcttccaa 20 <210> 1518 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1518 ttaaatgatc taactcttct 20 <210> 1519 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1519 tgagaaagtt aaatgatcta 20 <210> 1520 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1520 tacttaaatt tttagagttt 20 <210> 1521 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1521 aaagttactt aaatttttag 20 <210> 1522 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1522 atcttaaagt tacttaaatt 20 <210> 1523 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1523 atgtgatctt aaagttactt 20 <210> 1524 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1524 taactatgtg atcttaaagt 20 <210> 1525 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1525 ttactctttt ctactaagta 20 <210> 1526 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1526 gggtattact cttttctact 20 <210> 1527 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1527 taaagtttgc ttgctgggta 20 <210> 1528 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1528 tattgtaaag tttgcttgct 20 <210> 1529 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1529 taaaaggatc tattgtaaag 20 <210> 1530 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1530 ttatttaaaa ggatctattg 20 <210> 1531 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1531 ggaccttatt taaaaggatc 20 <210> 1532 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1532 gatatttcct aggaccttat 20 <210> 1533 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1533 tgaatgatat ttcctaggac 20 <210> 1534 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1534 gatgctggca tgaatgatat 20 <210> 1535 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1535 ttttttgatg ctggcatgaa 20 <210> 1536 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1536 gttagttttt tgatgctggc 20 <210> 1537 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1537 ttagtgttag ttttttgatg 20 <210> 1538 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1538 gcattattag tgttagtttt 20 <210> 1539 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1539 atcttgcatt attagtgtta 20 <210> 1540 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1540 ataatatctt gcattattag 20 <210> 1541 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1541 cagtaagaaa agcagaatat 20 <210> 1542 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1542 tcattgacag taagaaaagc 20 <210> 1543 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1543 gatagttttt ctcattgaca 20 <210> 1544 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1544 gtttgcaatt tattgaatga 20 <210> 1545 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1545 gtgttgggtt tgcaatttat 20 <210> 1546 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1546 ttaagtgtgt tgggtttgca 20 <210> 1547 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1547 ttttatttaa gtgtgttggg 20 <210> 1548 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1548 tttagcagta acattttatt 20 <210> 1549 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1549 gttagtttag cagtaacatt 20 <210> 1550 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1550 tctatatatt cagtagttta 20 <210> 1551 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1551 ttactttcta tatattcagt 20 <210> 1552 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1552 gtttgcttac tttctata 20 <210> 1553 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1553 agtttgtttg cttactttct 20 <210> 1554 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1554 tggcaagttt gtttgcttac 20 <210> 1555 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1555 ttactgttac tgtatttccc 20 <210> 1556 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1556 atgtagttac tgttactgta 20 <210> 1557 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1557 atgcaattta atgggtacag 20 <210> 1558 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1558 tagatatgca atttaatggg 20 <210> 1559 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1559 aggagataga tatgcaattt 20 <210> 1560 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1560 cctaaaggag atagatatgc 20 <210> 1561 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1561 agcctcctaa aggagataga 20 <210> 1562 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1562 caccaccagc ctcctaaagg 20 <210> 1563 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1563 atctaagaaa attaataaac 20 <210> 1564 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1564 atgatcacat ctaagaaaat 20 <210> 1565 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1565 aactgcaata ccatgatcac 20 <210> 1566 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1566 taaaactgca ataccatgat 20 <210> 1567 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1567 ctttaaaact gcaataccat 20 <210> 1568 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1568 tctcctttaa aactgcaata 20 <210> 1569 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1569 tgttctcctt taaaactgca 20 <210> 1570 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1570 gattgttctc ctttaaaact 20 <210> 1571 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1571 aggagatgt tctcctttaa 20 <210> 1572 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1572 aacaggagat tgttctcctt 20 <210> 1573 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1573 ttaaacagga gattgttctc 20 <210> 1574 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1574 ctcttaaaca ggagattgtt 20 <210> 1575 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1575 ctttaactcc gtaaatattt 20 <210> 1576 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1576 agtgaccttt aactccgtaa 20 <210> 1577 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1577 ggagtccagt gacctttaac 20 <210> 1578 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1578 accagtctgg agtccagtga 20 <210> 1579 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1579 tcatcttacc aaactatttt 20 <210> 1580 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1580 gaatcatctt accaaactat 20 <210> 1581 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1581 taagaatcat cttaccaaac 20 <210> 1582 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1582 aagaatgtaa gaatcatctt 20 <210> 1583 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1583 gttatttaag aatgtaagaa 20 <210> 1584 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1584 cgtgttattt aagaatgtaa 20 <210> 1585 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1585 taaagcattt ttcttagatg 20 <210> 1586 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1586 tgttaaagca tttttcttag 20 <210> 1587 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1587 tttatgttaa agcatttttc 20 <210> 1588 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1588 atgtttatgt taaagcattt 20 <210> 1589 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1589 gcattttttc agtaatgttt 20 <210> 1590 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1590 tgtagcattt tttcagtaat 20 <210> 1591 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1591 caaatgtagc attttttcag 20 <210> 1592 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1592 aagttgtggc aaatgtagca 20 <210> 1593 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1593 atgaagttgt ggcaaatgta 20 <210> 1594 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1594 tttatgaagt tgtggcaaat 20 <210> 1595 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1595 tgacatttta tgaagttgtg 20 <210> 1596 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1596 cttgagattt cacttgacat 20 <210> 1597 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1597 tttggagctt gagattcac 20 <210> 1598 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1598 atctttggag cttgagattt 20 <210> 1599 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1599 aatatctttg gagcttgaga 20 <210> 1600 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1600 aataatatct ttggagcttg 20 <210> 1601 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1601 aggaataata tctttggagc 20 <210> 1602 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1602 aataggaata atatctttgg 20 <210> 1603 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1603 agtaatagga ataatatctt 20 <210> 1604 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1604 ttacatcaga tttagtaata 20 <210> 1605 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1605 aaatgttatt acatcagatt 20 <210> 1606 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1606 ataaaatgtt attacatcag 20 <210> 1607 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1607 cctagaatca ataaaatgtt 20 <210> 1608 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1608 aggaatgcct agaatcaata 20 <210> 1609 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1609 attcagcagg aatgcctaga 20 <210> 1610 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1610 tacattcagc aggaatgcct 20 <210> 1611 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1611 ttacctgata taacatcaca 20 <210> 1612 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1612 gttttacctg atataacatc 20 <210> 1613 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1613 caggttttac ctgatataac 20 <210> 1614 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1614 ttagacaggt tttacctgat 20 <210> 1615 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1615 attctcctta gacaggtttt 20 <210> 1616 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1616 actgtctatt ctccttagac 20 <210> 1617 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1617 actactgtct attctcctta 20 <210> 1618 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1618 actaactact gtctattctc 20 <210> 1619 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1619 tgaactaact actgtctatt 20 <210> 1620 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1620 agttgaacta actactgtct 20 <210> 1621 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1621 attaattgat atgtaaaacg 20 <210> 1622 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1622 ccaattaatt gatatgtaaa 20 <210> 1623 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1623 tccttttaac ttccaattaa 20 <210> 1624 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1624 gtttcctggt ccttttaact 20 <210> 1625 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1625 tctgagtttc ctggtccttt 20 <210> 1626 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1626 atgtctgagt ttcctggtcc 20 <210> 1627 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1627 tgtatgtctg agtttcctgg 20 <210> 1628 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1628 atgtatactg tatgtctgag 20 <210> 1629 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1629 aaaatgtata ctgtatgtct 20 <210> 1630 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1630 ttttaaaatg tatactgtat 20 <210> 1631 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1631 catacattct atatattata 20 <210> 1632 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1632 attataagcc atacattcta 20 <210> 1633 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1633 ttcattataa gccatacatt 20 <210> 1634 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1634 taattcatta taagccatac 20 <210> 1635 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1635 tgagttaact aattcattat 20 <210> 1636 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1636 tttgcattga gttaactaat 20 <210> 1637 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1637 taatttgcat tgagttaact 20 <210> 1638 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1638 gaataatttg cattgagtta 20 <210> 1639 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1639 atagaataat ttgcattgag 20 <210> 1640 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1640 aaaatagaat aatttgcatt 20 <210> 1641 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1641 ttgtaatcaa aatagaataa 20 <210> 1642 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1642 tatttgtaat caaaatagaa 20 <210> 1643 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1643 tactatttgt aatcaaaata 20 <210> 1644 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1644 ttttactatt tgtaatcaaa 20 <210> 1645 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1645 gcttatttta ctatttgtaa 20 <210> 1646 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1646 cttgcttatt ttactatttg 20 <210> 1647 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1647 ttatcttgct tatttacta 20 <210> 1648 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1648 gttattttat cttgcttatt 20 <210> 1649 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1649 aaacatctgt tattttatct 20 <210> 1650 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1650 ggattttaaa catctgttat 20 <210> 1651 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1651 ctttttggat tttaaacatc 20 <210> 1652 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1652 gtgctttttg gattttaaac 20 <210> 1653 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1653 ttttgtatgt gctttttgga 20 <210> 1654 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1654 taagtgagta cttagacatc 20 <210> 1655 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1655 tactttataa gtgagtactt 20 <210> 1656 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1656 ttctacttta taagtgagta 20 <210> 1657 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1657 aatgtcttct actttataag 20 <210> 1658 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1658 aataatgaat gtcttctact 20 <210> 1659 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1659 tataataatg aatgtcttct 20 <210> 1660 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1660 tgatataata atgaatgtct 20 <210> 1661 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1661 aaaatttgat ataataatga 20 <210> 1662 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1662 catttaaaaa tttgatataa 20 <210> 1663 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1663 gtactgagca tttaaaaatt 20 <210> 1664 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1664 aatggtcaaa tagtactgag 20 <210> 1665 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1665 ttaaatggtc aaatagtact 20 <210> 1666 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1666 agtttgaata caaaattttt 20 <210> 1667 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1667 actggtagtt tgaatacaaa 20 <210> 1668 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1668 ttcactggta gtttgaatac 20 <210> 1669 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1669 gctttcactg gtagtttgaa 20 <210> 1670 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1670 agggctttca ctggtagttt 20 <210> 1671 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1671 ggtagggctt tcactggtag 20 <210> 1672 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1672 ctaggtaggg ctttcactgg 20 <210> 1673 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1673 cttctaggta gggctttcac 20 <210> 1674 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1674 taccttctag gtagggcttt 20 <210> 1675 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1675 gtataccttc taggtagggc 20 <210> 1676 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1676 tgagtatacc ttctaggtag 20 <210> 1677 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1677 cactgagtat accttctagg 20 <210> 1678 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1678 tatcactgag tataccttct 20 <210> 1679 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1679 acttatcact gagtatacct 20 <210> 1680 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1680 acaaaactta tcactgagta 20 <210> 1681 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1681 gctacaaaac ttatcactga 20 <210> 1682 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1682 ggagctacaa aacttatcac 20 <210> 1683 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1683 gatttggagc tacaaaactt 20 <210> 1684 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1684 gaagattgg agctacaaaa 20 <210> 1685 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1685 ctattagaag atttggagct 20 <210> 1686 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1686 cactcactat tagaagattt 20 <210> 1687 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1687 tgtcagcctt ttattttggg 20 <210> 1688 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1688 acctgtcagc cttttatttt 20 <210> 1689 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1689 tcgacttacc tgtcagcctt 20 <210> 1690 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1690 ttctcgactt acctgtcagc 20 <210> 1691 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1691 gtattctcga cttacctgtc 20 <210> 1692 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1692 taacatccat atacagtcaa 20 <210> 1693 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1693 tattaacatc catatacagt 20 <210> 1694 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1694 atttattaac atccatatac 20 <210> 1695 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1695 gctatttatt aacatccata 20 <210> 1696 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1696 ctgtcagcta tttattaaca 20 <210> 1697 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1697 ttactgtcag ctattttatta 20 <210> 1698 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1698 actttactgt cagctattta 20 <210> 1699 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1699 taaactttac tgtcagctat 20 <210> 1700 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1700 ggataaactt tactgtcagc 20 <210> 1701 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1701 tatggataaa ctttactgtc 20 <210> 1702 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1702 ttatatggat aaactttact 20 <210> 1703 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1703 ttgcaagtct ttatatggat 20 <210> 1704 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1704 tatttgcaag tctttatatg 20 <210> 1705 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1705 gaatatttgc aagtctttat 20 <210> 1706 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1706 gtagaggaat atttgcaagt 20 <210> 1707 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1707 gttacattat tatagatatt 20 <210> 1708 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1708 tgtgttacat tattatagat 20 <210> 1709 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1709 gaaatgtgtt acattattat 20 <210> 1710 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1710 ttcaccagtg aaatgtgtta 20 <210> 1711 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1711 tgtttcacca gtgaaatgtg 20 <210> 1712 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1712 acatgtttca ccagtgaaat 20 <210> 1713 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1713 aagacatgtt tcaccagtga 20 <210> 1714 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1714 gacaagacat gtttcaccag 20 <210> 1715 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1715 tatgacaaga catgtttcac 20 <210> 1716 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1716 gcatatgaca agacatgttt 20 <210> 1717 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1717 taatgcatat gacaagacat 20 <210> 1718 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1718 ctataatgca tatgacaaga 20 <210> 1719 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1719 tttctataat gcatatgaca 20 <210> 1720 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1720 tcctttctat aatgcatatg 20 <210> 1721 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1721 tctgattatc ctttctataa 20 <210> 1722 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1722 aagtctgatt atcctttcta 20 <210> 1723 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1723 aactgaaagt ctgattatcc 20 <210> 1724 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1724 tataactgaa agtctgatta 20 <210> 1725 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1725 gttaaaaata ttaatataac 20 <210> 1726 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1726 tgtgcacaaa aatgttaaaa 20 <210> 1727 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1727 ctatgtgcac aaaaatgtta 20 <210> 1728 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1728 tagctatgtg cacaaaaatg 20 <210> 1729 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1729 agatagctat gtgcacaaaa 20 <210> 1730 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1730 tgaagatagc tatgtgcaca 20 <210> 1731 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1731 tattgaagat agctatgtgc 20 <210> 1732 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1732 ttttattgaa gatagctatg 20 <210> 1733 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1733 caattttatt gaagatagct 20 <210> 1734 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1734 aaacaatttt attgaagata 20 <210> 1735 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1735 gtgtatctta aaataatacc 20 <210> 1736 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1736 ttagtgtatc ttaaaataat 20 <210> 1737 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1737 tgatcatttt agtgtatctt 20 <210> 1738 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1738 cccttgatca ttttagtgta 20 <210> 1739 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1739 aatcccttga tcattttagt 20 <210> 1740 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1740 ttgaatccct tgatcatttt 20 <210> 1741 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1741 ttagtcttga atcccttgat 20 <210> 1742 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1742 ttgtttagtc ttgaatccct 20 <210> 1743 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1743 gagttgttta gtcttgaatc 20 <210> 1744 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1744 attgagttgt ttagtcttga 20 <210> 1745 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1745 ctaattgagt tgtttagtct 20 <210> 1746 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1746 caactaattg agttgtttag 20 <210> 1747 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1747 attggtgcaa ctaattgagt 20 <210> 1748 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1748 tttattggtg caactaattg 20 <210> 1749 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1749 ttttttattg gtgcaactaa 20 <210> 1750 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1750 taagtgtttt ttattggtgc 20 <210> 1751 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1751 actgacagtt tttttaagtg 20 <210> 1752 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1752 gacactgaca gtttttttaa 20 <210> 1753 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1753 ttggacactg acagtttttt 20 <210> 1754 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1754 aggttggaca ctgacagttt 20 <210> 1755 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1755 tacaggttgg acactgacag 20 <210> 1756 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1756 tatattggat aatttgaaat 20 <210> 1757 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1757 atgtatattg gataatttga 20 <210> 1758 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1758 gacatgtata ttggataatt 20 <210> 1759 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1759 atgacatgta tattggataa 20 <210> 1760 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1760 tatatatgac atgtatattg 20 <210> 1761 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1761 atgtgacata taaaaatata 20 <210> 1762 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1762 atatgtgaca tataaaaata 20 <210> 1763 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1763 tttatatatg tgacatataa 20 <210> 1764 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1764 cttttatata tgtgacatat 20 <210> 1765 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1765 atcttttata tatgtgacat 20 <210> 1766 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1766 catatctttt atatatgtga 20 <210> 1767 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1767 tcatacatat cttttatata 20 <210> 1768 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1768 tagatcatac atatctttta 20 <210> 1769 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1769 catagatcat acatatcttt 20 <210> 1770 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1770 cacatagatc atacatatct 20 <210> 1771 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1771 aggattcaca tagatcatac 20 <210> 1772 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1772 ttaggattca catagatcat 20 <210> 1773 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1773 acttaggatt cacatagatc 20 <210> 1774 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1774 ttacttagga ttcacataga 20 <210> 1775 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1775 tatttactta ggattcacat 20 <210> 1776 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1776 aatatttact taggattcac 20 <210> 1777 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1777 tgtacttttc tggaacaaaa 20 <210> 1778 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1778 gattattttt acctttatta 20 <210> 1779 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1779 tagattattt ttacctttat 20 <210> 1780 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1780 attatagatt atttttacct 20 <210> 1781 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1781 gaaaattata gattattttt 20 <210> 1782 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1782 ggtcctgaaa attatagatt 20 <210> 1783 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1783 gtggtcctga aaattataga 20 <210> 1784 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1784 ctgtggtcct gaaaattata 20 <210> 1785 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1785 gtctgtggtc ctgaaaatta 20 <210> 1786 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1786 tcgacagctt agtctgtggt 20 <210> 1787 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1787 tttcgacagc ttagtctgtg 20 <210> 1788 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1788 aatttcgaca gcttagtctg 20 <210> 1789 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1789 ttaatttcga cagcttagtc 20 <210> 1790 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1790 cgttaatttc gacagcttag 20 <210> 1791 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1791 tggccctaaa aaaatcagcg 20 <210> 1792 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1792 tctggcccta aaaaaatcag 20 <210> 1793 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1793 tggtattctg gccctaaaaa 20 <210> 1794 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1794 tttggtattc tggccctaaa 20 <210> 1795 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1795 ccattttggt attctggccc 20 <210> 1796 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1796 gaggagccat tttggtattc 20 <210> 1797 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1797 gagaggagcc attttggtat 20 <210> 1798 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1798 aagagaggag ccattttggt 20 <210> 1799 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1799 tgaaattgtc caattttggg 20 <210> 1800 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1800 tttgaaattg tccaattttg 20 <210> 1801 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1801 catttgaaat tgtccaattt 20 <210> 1802 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1802 tgcatttgaa attgtccaat 20 <210> 1803 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1803 attttgcatt tgaaattgtc 20 <210> 1804 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1804 ataatgaatt attttgcatt 20 <210> 1805 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1805 taaataatga attattttgc 20 <210> 1806 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1806 ctcatatatt aaataatgaa 20 <210> 1807 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1807 aactcatata ttaaataatg 20 <210> 1808 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1808 tagaggaagc aactcatata 20 <210> 1809 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1809 aatagaggaa gcaactcata 20 <210> 1810 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1810 caaatagagg aagcaactca 20 <210> 1811 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1811 accaaataga ggaagcaact 20 <210> 1812 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1812 aaaccaaata gaggaagcaa 20 <210> 1813 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1813 ggaaaccaaa tagaggaagc 20 <210> 1814 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1814 taaggaaacc aaatagagga 20 <210> 1815 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1815 tttaaggaaa ccaaatagag 20 <210> 1816 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1816 tgttttcttc tggaagcaga 20 <210> 1817 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1817 cttactttaa gtgaagttac 20 <210> 1818 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1818 ttttctactt actttaagtg 20 <210> 1819 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1819 acatgaaccc tctttatttt 20 <210> 1820 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1820 gaaaacataa acatgaaccc 20 <210> 1821 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1821 agatccacat tgaaaacata 20 <210> 1822 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1822 ttaaaagatc cacattgaaa 20 <210> 1823 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1823 gccttagaaa tatttttttt 20 <210> 1824 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1824 caaatggcat gccttagaaa 20 <210> 1825 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1825 tatttcaaat ggcatgcctt 20 <210> 1826 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1826 caaagtattt caaatggcat 20 <210> 1827 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1827 tgcaacaaag tatttcaaat 20 <210> 1828 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1828 tcaacaatgc aacaaagtat 20 <210> 1829 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1829 gaaaaaaaag tatttcaaca 20 <210> 1830 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1830 gattattttt cttggaaaaa 20 <210> 1831 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1831 gaaattttat tttctggaga 20 <210> 1832 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1832 aaattataat aggaaatttt 20 <210> 1833 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1833 ctgaatataa tgaatgaaat 20 <210> 1834 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1834 tacctgaata taatgaatga 20 <210> 1835 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1835 gactacctga atataatgaa 20 <210> 1836 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1836 atggactacc tgaatataat 20 <210> 1837 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1837 tccatggact acctgaatat 20 <210> 1838 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1838 accatcaagc ctcccaaaac 20 <210> 1839 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1839 ccttaccatc aagcctccca 20 <210> 1840 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1840 agtcccctta ccatcaagcc 20 <210> 1841 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1841 tgtagtcccc taccatcaa 20 <210> 1842 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1842 gaatgtagtc cccttaccat 20 <210> 1843 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1843 attgaatgta gtccccttac 20 <210> 1844 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1844 atgattgaat gtagtcccct 20 <210> 1845 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1845 gattagcaag tgaatgaatg 20 <210> 1846 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1846 gtagattagc aagtgaatga 20 <210> 1847 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1847 tttgtagatt agcaagtgaa 20 <210> 1848 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1848 atatttgtag attagcaagt 20 <210> 1849 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1849 ccataagagg ttctcagtaa 20 <210> 1850 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1850 ggtccataag aggttctcag 20 <210> 1851 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1851 cctggtccat aagaggttct 20 <210> 1852 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1852 taatacctgg tccataagag 20 <210> 1853 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1853 tcctaatacc tggtccataa 20 <210> 1854 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1854 ttttcctaat acctggtcca 20 <210> 1855 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1855 tacttttcct aatacctggt 20 <210> 1856 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1856 cgttactact tttcctaata 20 <210> 1857 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1857 aaggctgaga ctgcttctcg 20 <210> 1858 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1858 gataataaat tatatgaagg 20 <210> 1859 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1859 gtttgataat aaattatatg 20 <210> 1860 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1860 gtgtaattgt ttgataataa 20 <210> 1861 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1861 aatgtgtaat tgtttgataa 20 <210> 1862 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1862 gtaatttact aacaaatgtg 20 <210> 1863 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1863 agtgtaattt actaacaaat 20 <210> 1864 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1864 ataagtgtaa tttaactaaca 20 <210> 1865 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1865 gtaataagtg taatttacta 20 <210> 1866 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1866 gttgtaataa gtgtaattta 20 <210> 1867 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1867 acagttgtaa taagtgtaat 20 <210> 1868 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1868 ataacagttg taataagtgt 20 <210> 1869 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1869 ttcaaataat aacagttgta 20 <210> 1870 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1870 ataattcaaa taataacagt 20 <210> 1871 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1871 aattgtgata aatataattc 20 <210> 1872 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1872 atgtaattgt gataaatata 20 <210> 1873 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1873 gacatgtaat tgtgataaat 20 <210> 1874 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1874 acagacatgt aattgtgata 20 <210> 1875 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1875 agaacagaca tgtaattgtg 20 <210> 1876 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1876 ttaagaacag acatgtaatt 20 <210> 1877 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1877 aagtatattt aagaacagac 20 <210> 1878 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1878 ttaaattgtg ataagtatat 20 <210> 1879 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1879 gaattaaatt gtgataagta 20 <210> 1880 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1880 gtggaattaa attgtgataa 20 <210> 1881 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1881 gccgtggaat taaattgtga 20 <210> 1882 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1882 taagccgtgg aattaaattg 20 <210> 1883 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1883 ttgtaagccg tggaattaaa 20 <210> 1884 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1884 tcattgtaag ccgtggaatt 20 <210> 1885 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1885 tgatcattgt aagccgtgga 20 <210> 1886 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1886 tatagttatg atcattgtaa 20 <210> 1887 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1887 aattatagtt atgatcattg 20 <210> 1888 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1888 ctttaataat tatagttatg 20 <210> 1889 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1889 tgtctttaat aattatagtt 20 <210> 1890 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1890 aattgtcttt aataattata 20 <210> 1891 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1891 tcaaaattgt ctttaataat 20 <210> 1892 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1892 atttaatcaa aattgtcttt 20 <210> 1893 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1893 taacatttaa tcaaaattgt 20 <210> 1894 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1894 acataacatt taatcaaaat 20 <210> 1895 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1895 atgacataac atttaatcaa 20 <210> 1896 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1896 tacttatgac ataacattta 20 <210> 1897 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1897 ttactactta tgacataaca 20 <210> 1898 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1898 aacagttact acttatgaca 20 <210> 1899 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1899 tgtaacagtt actacttatg 20 <210> 1900 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1900 cttatttgta acagttacta 20 <210> 1901 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1901 tttcacagct tatttgtaac 20 <210> 1902 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1902 tcttttcaca gcttatttgt 20 <210> 1903 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1903 ggttcttttc acagcttatt 20 <210> 1904 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1904 ctaggagtgg ttcttttcac 20 <210> 1905 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1905 atgctaggag tggttctttt 20 <210> 1906 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1906 ctaatgctag gagtggttct 20 <210> 1907 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1907 agagtgacta atgctaggag 20 <210> 1908 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1908 agagaataga gtgactaatg 20 <210> 1909 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1909 ttaatgagag aatagagtga 20 <210> 1910 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1910 ctgttggttt aattgtttat 20 <210> 1911 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1911 tgctgttggt ttaattgttt 20 <210> 1912 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1912 tatgctgttg gtttaattgt 20 <210> 1913 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1913 actatgctgt tggtttaatt 20 <210> 1914 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1914 tgactatgct gttggtttaa 20 <210> 1915 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1915 atttgactat gctgttggtt 20 <210> 1916 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1916 ttatttgact atgctgttgg 20 <210> 1917 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1917 ttttatttga ctatgctgtt 20 <210> 1918 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1918 tcttttattt gactatgctg 20 <210> 1919 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1919 tttcttttat ttgactatgc 20 <210> 1920 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1920 cttctgagct gattttctat 20 <210> 1921 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1921 tccttctgag ctgattttct 20 <210> 1922 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1922 agtccttctg agctgatttt 20 <210> 1923 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1923 ctagtccttc tgagctgatt 20 <210> 1924 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1924 tactagtcct tctgagctga 20 <210> 1925 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1925 aatactagtc cttctgagct 20 <210> 1926 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1926 tgaatactag tccttctgag 20 <210> 1927 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1927 cttgaatact agtccttctg 20 <210> 1928 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1928 ttcttgaata ctagtccttc 20 <210> 1929 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1929 tgggttcttg aatactagtc 20 <210> 1930 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1930 tgtgggttct tgaatactag 20 <210> 1931 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1931 tctgtgggtt cttgaatact 20 <210> 1932 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1932 tagagaaatt tctgtgggtt 20 <210> 1933 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1933 aagatagaga aatttctgtg 20 <210> 1934 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1934 ggaagataga gaaatttctg 20 <210> 1935 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1935 cttggcttgg aagatagaga 20 <210> 1936 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1936 ctcttggctt ggaagataga 20 <210> 1937 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1937 aagggagtag ttcttggtgc 20 <210> 1938 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1938 aaagggagta gttcttggtg 20 <210> 1939 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1939 gaaagggagt agttcttggt 20 <210> 1940 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1940 agaaagggag tagttcttgg 20 <210> 1941 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1941 aagaaaggga gtagttcttg 20 <210> 1942 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1942 gaagaaaggg agtagttctt 20 <210> 1943 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1943 tcaactgaag aaagggagta 20 <210> 1944 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1944 attcaactga agaaagggag 20 <210> 1945 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1945 cattcaactg aagaaaggga 20 <210> 1946 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1946 tcattcaact gaagaaaggg 20 <210> 1947 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1947 tttcattcaa ctgaagaaag 20 <210> 1948 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1948 atttcattca actgaagaaa 20 <210> 1949 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1949 tatttcattc aactgaagaa 20 <210> 1950 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1950 ttatttcatt caactgaaga 20 <210> 1951 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1951 cttatttcat tcaactgaag 20 <210> 1952 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1952 tcttatttca ttcaactgaa 20 <210> 1953 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1953 ttcttatttc attcaactga 20 <210> 1954 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1954 atttcttatt tcattcaact 20 <210> 1955 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1955 tacattctt atttcattca 20 <210> 1956 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1956 ttcagcagga atgccatcat 20 <210> 1957 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1957 attcagcagg aatgccatca 20 <210> 1958 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1958 cattcagcag gaatgccatc 20 <210> 1959 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1959 acattcagca ggaatgccat 20 <210> 1960 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1960 tacattcagc aggaatgcca 20 <210> 1961 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1961 gtacattcag caggaatgcc 20 <210> 1962 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1962 tggtacattc agcaggaatg 20 <210> 1963 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1963 gtggtacatt cagcaggaat 20 <210> 1964 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1964 ggtggtacat tcagcaggaa 20 <210> 1965 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1965 tggtggtaca ttcagcagga 20 <210> 1966 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1966 atggtggtac attcagcagg 20 <210> 1967 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1967 aatggtggta cattcagcag 20 <210> 1968 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1968 aaatggtggt acatcagca 20 <210> 1969 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1969 taaatggtgg tacattcagc 20 <210> 1970 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1970 tataaatggt ggtacattca 20 <210> 1971 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1971 ttataaatgg tggtacattc 20 <210> 1972 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1972 gttataaatg gtggtacatt 20 <210> 1973 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1973 tgttataaat ggtggtacat 20 <210> 1974 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1974 ctgttataaa tggtggtaca 20 <210> 1975 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1975 tctgttataa atggtggtac 20 <210> 1976 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1976 cacctctgtt ataaatggtg 20 <210> 1977 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1977 tcacctctgt tataaatggt 20 <210> 1978 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1978 ttcacctctg ttataaatgg 20 <210> 1979 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1979 gttcacctct gttataaatg 20 <210> 1980 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1980 atgttcacct ctgttataaa 20 <210> 1981 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1981 tatgttcacc tctgttataa 20 <210> 1982 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1982 gtatgttcac ctctgttata 20 <210> 1983 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1983 tgtatgttca cctctgttat 20 <210> 1984 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1984 taatcgcaac tagatgtagc 20 <210> 1985 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1985 gtaatcgcaa ctagatgtag 20 <210> 1986 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1986 agtaatcgca actagatgta 20 <210> 1987 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1987 cagtaatcgc aactagatgt 20 <210> 1988 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1988 ccagtaatcg caactagatg 20 <210> 1989 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1989 gccagtaatc gcaactagat 20 <210> 1990 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1990 tgccagtaat cgcaactaga 20 <210> 1991 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1991 ttgccagtaa tcgcaactag 20 <210> 1992 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1992 attgccagta atcgcaacta 20 <210> 1993 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1993 cattgccagt aatcgcaact 20 <210> 1994 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1994 acattgccag taatcgcaac 20 <210> 1995 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1995 gacattgcca gtaatcgcaa 20 <210> 1996 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1996 gggacattgc cagtaatcgc 20 <210> 1997 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1997 ttgttttccg ggattgcatt 20 <210> 1998 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1998 tttgttttcc gggattgcat 20 <210> 1999 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 1999 aatctttgtt ttccgggatt 20 <210> 2000 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2000 aaatctttgt tttccgggat 20 <210> 2001 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2001 aaacaccaaa tctttgtttt 20 <210> 2002 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2002 aaaacaccaa atctttgttt 20 <210> 2003 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2003 gtagaaaaca ccaaatcttt 20 <210> 2004 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2004 agtagaaaac accaaatctt 20 <210> 2005 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2005 cccaagtaga aaacaccaaa 20 <210> 2006 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2006 tcccaagtag aaaacaccaa 20 <210> 2007 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2007 gtgatcccaa gtagaaaaca 20 <210> 2008 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2008 tgtgatccca agtagaaaac 20 <210> 2009 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2009 ttgtgatccc aagtagaaaa 20 <210> 2010 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2010 tttgtgatcc caagtagaaa 20 <210> 2011 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2011 cttttgcttt gtgatcccaa 20 <210> 2012 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2012 tgtccttttg ctttgtgatc 20 <210> 2013 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2013 gtgtcctttt gctttgtgat 20 <210> 2014 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2014 agtgtccttt tgctttgtga 20 <210> 2015 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2015 ttgaagtgtc cttttgcttt 20 <210> 2016 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2016 gttgaagtgt ccttttgctt 20 <210> 2017 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2017 agttgaagtg tccttttgct 20 <210> 2018 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2018 cagttgaagt gtccttttgc 20 <210> 2019 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2019 acagttgaag tgtccttttg 20 <210> 2020 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2020 gacagttgaa gtgtcctttt 20 <210> 2021 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2021 ggacagttga agtgtccttt 20 <210> 2022 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2022 tggacagttg aagtgtcctt 20 <210> 2023 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2023 ctggacagtt gaagtgtcct 20 <210> 2024 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2024 tctggacagt tgaagtgtcc 20 <210> 2025 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2025 ctctggacag ttgaagtgtc 20 <210> 2026 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2026 cctctggaca gttgaagtgt 20 <210> 2027 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2027 ccctctggac agttgaagtg 20 <210> 2028 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2028 accctctgga cagttgaagt 20 <210> 2029 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2029 aaccctctgg acagttgaag 20 <210> 2030 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2030 taaccctctg gacagttgaa 20 <210> 2031 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2031 ataaccctct ggacagttga 20 <210> 2032 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2032 aataaccctc tggacagttg 20 <210> 2033 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2033 gaataaccct ctggacagtt 20 <210> 2034 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2034 tgaataaccc tctggacagt 20 <210> 2035 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2035 ctgaataacc ctctggacag 20 <210> 2036 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2036 cctgaataac cctctggaca 20 <210> 2037 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2037 tcctgaataa ccctctggac 20 <210> 2038 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2038 ctcctgaata accctctgga 20 <210> 2039 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2039 cctcctgaat aaccctctgg 20 <210> 2040 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2040 gcctcctgaa taaccctctg 20 <210> 2041 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2041 agcctcctga ataaccctct 20 <210> 2042 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2042 cagcctcctg aataaccctc 20 <210> 2043 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2043 ccagcctcct gaataaccct 20 <210> 2044 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2044 accagcctcc tgaataaccc 20 <210> 2045 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2045 caccagcctc ctgaataacc 20 <210> 2046 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2046 ccaccagcct cctgaataac 20 <210> 2047 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2047 accaccagcc tcctgaataa 20 <210> 2048 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2048 caccaccagc ctcctgaata 20 <210> 2049 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2049 ccaccaccag cctcctgaat 20 <210> 2050 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2050 catgccacca ccagcctcct 20 <210> 2051 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2051 atcatgccac caccagcctc 20 <210> 2052 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2052 catcatgcca ccaccagcct 20 <210> 2053 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2053 acactcatca tgccaccacc 20 <210> 2054 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2054 ttctccacac tcatcatgcc 20 <210> 2055 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2055 ttttctccac actcatcatg 20 <210> 2056 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2056 gttttctcca cactcatcat 20 <210> 2057 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2057 atttaagaac tgtacaatta 20 <210> 2058 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2058 catttaagaa ctgtacaatt 20 <210> 2059 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2059 acatttaaga actgtacaat 20 <210> 2060 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2060 aacattaag aactgtacaa 20 <210> 2061 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2061 caacatttaa gaactgtaca 20 <210> 2062 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2062 acaacattta agaactgtac 20 <210> 2063 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2063 tacaacattt aagaactgta 20 <210> 2064 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2064 ctacaactt taagaactgt 20 <210> 2065 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2065 actacaacat ttaagaactg 20 <210> 2066 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2066 tactacaaca tttaagaact 20 <210> 2067 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2067 atactacaac atttaagaac 20 <210> 2068 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2068 aatactacaa catttaagaa 20 <210> 2069 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2069 taatactaca acatttaaga 20 <210> 2070 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2070 ttaatactac aacattaag 20 <210> 2071 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2071 gaaattaata ctacaacatt 20 <210> 2072 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2072 ttttgaaatt aatactacaa 20 <210> 2073 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2073 gttttgaaat taatactaca 20 <210> 2074 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2074 agttttgaaa ttaatactac 20 <210> 2075 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2075 tagttttgaa attaatacta 20 <210> 2076 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2076 ttagttttga aattatact 20 <210> 2077 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2077 cgatttttag ttttgaaatt 20 <210> 2078 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2078 acgattttta gttttgaaat 20 <210> 2079 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2079 gacgattttt agttttgaaa 20 <210> 2080 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2080 tgacgatttt tagttttgaa 20 <210> 2081 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2081 ctgacgattt ttagttttga 20 <210> 2082 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2082 tgctgacgat ttttagtttt 20 <210> 2083 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2083 gtgctgacga tttttagttt 20 <210> 2084 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2084 tgtgctgacg atttttagtt 20 <210> 2085 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2085 ctgtgctgac gatttttagt 20 <210> 2086 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2086 tctgtgctga cgatttttag 20 <210> 2087 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2087 ctctgtgctg acgattttta 20 <210> 2088 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2088 actctgtgct gacgattttt 20 <210> 2089 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2089 tactctgtgc tgacgatttt 20 <210> 2090 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2090 atactctgtg ctgacgattt 20 <210> 2091 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2091 catactctgt gctgacgatt 20 <210> 2092 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2092 acatactctg tgctgacgat 20 <210> 2093 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2093 tttttacaca tactctgtgc 20 <210> 2094 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2094 cagatttta cacatactct 20 <210> 2095 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2095 acagatttt acacatactc 20 <210> 2096 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2096 tacagattt tacacatact 20 <210> 2097 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2097 ttacagattt ttacacatac 20 <210> 2098 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2098 tattacagat ttttacacat 20 <210> 2099 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2099 tgtattacag atttttacac 20 <210> 2100 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2100 cagtttaaaa atttgtatta 20 <210> 2101 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2101 catcagttta aaaatttgta 20 <210> 2102 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2102 aagcatcagt ttaaaaattt 20 <210> 2103 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2103 gaagcatcag tttaaaaatt 20 <210> 2104 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2104 tagcaaaatg aagcatcagt 20 <210> 2105 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2105 gtagcaaaat gaagcatcag 20 <210> 2106 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2106 tgtagcaaaa tgaagcatca 20 <210> 2107 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2107 ttgtagcaaa atgaagcatc 20 <210> 2108 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2108 tttgtagcaa aatgaagcat 20 <210> 2109 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2109 aacatttact ccaaattatt 20 <210> 2110 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2110 caaacattta ctccaaatta 20 <210> 2111 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2111 atcaaacatt tactccaaat 20 <210> 2112 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2112 catatcaaac atttactcca 20 <210> 2113 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2113 tcatatcaaa catttactcc 20 <210> 2114 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2114 atcatatcaa acatttactc 20 <210> 2115 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2115 taaataaatc atatcaaaca 20 <210> 2116 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2116 tcataaataa atcatatcaa 20 <210> 2117 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2117 ttcataaata aatcatatca 20 <210> 2118 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2118 tttcataaat aaatcatatc 20 <210> 2119 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2119 gtttcataaa taaatcatat 20 <210> 2120 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2120 ggtttcataa ataaatcata 20 <210> 2121 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2121 aggtttcata aataaatcat 20 <210> 2122 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2122 taggtttcat aaataaatca 20 <210> 2123 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2123 attaggtttc ataaataaat 20 <210> 2124 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2124 cattaggttt cataaataaa 20 <210> 2125 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2125 tcattaggtt tcataaataa 20 <210> 2126 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2126 ttcattaggt ttcataaata 20 <210> 2127 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2127 cttcattagg tttcataaat 20 <210> 2128 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2128 gcttcattag gtttcataaa 20 <210> 2129 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2129 ttctgcttca ttaggtttca 20 <210> 2130 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2130 taattctgct tcattaggtt 20 <210> 2131 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2131 tatgtagttc ttctcagttc 20 <210> 2132 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2132 tagtttatat gtagttcttc 20 <210> 2133 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2133 cttgtagttt atatgtagtt 20 <210> 2134 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2134 ttgacttgta gtttatatgt 20 <210> 2135 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2135 ttttgacttg tagtttatat 20 <210> 2136 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2136 atttttgact tgtagtttat 20 <210> 2137 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2137 tcttcatttt tgacttgtag 20 <210> 2138 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2138 tacctcttca tttttgactt 20 <210> 2139 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2139 attctttacc tcttcatttt 20 <210> 2140 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2140 caagtgacat attctttacc 20 <210> 2141 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2141 ttcaagtgac atattcttta 20 <210> 2142 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2142 ttgagttcaa gtgacatatt 20 <210> 2143 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2143 agttgagttc aagtgacata 20 <210> 2144 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2144 tttgagttga gttcaagtga 20 <210> 2145 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2145 tttcaagttt tgagttgagt 20 <210> 2146 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2146 gctttcaagt tttgagttga 20 <210> 2147 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2147 aggctttcaa gttttgagtt 20 <210> 2148 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2148 taggaggctt tcaagttttg 20 <210> 2149 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2149 ttctaggagg ctttcaagtt 20 <210> 2150 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2150 tcttctagga ggctttcaag 20 <210> 2151 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2151 gaattttttc ttctaggagg 20 <210> 2152 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2152 aagtagaatt ttttcttcta 20 <210> 2153 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2153 tgaagtagaa ttttttcttc 20 <210> 2154 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2154 gttgaagtag aattttttct 20 <210> 2155 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2155 tttgttgaag tagaattttt 20 <210> 2156 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2156 tttttgttga agtagaattt 20 <210> 2157 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2157 ttgctcttct aaatatttca 20 <210> 2158 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2158 agttgctctt ctaaatattt 20 <210> 2159 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2159 ttagttgctc ttctaaatat 20 <210> 2160 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2160 tagttagttg ctcttctaaa 20 <210> 2161 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2161 taagttagtt agttgctctt 20 <210> 2162 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2162 attaagttag ttagttgctc 20 <210> 2163 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2163 gaattaagtt agttagttgc 20 <210> 2164 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2164 ttgaattaag ttagttagtt 20 <210> 2165 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2165 ttttgaatta agttagttag 20 <210> 2166 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2166 tgattttgaa ttaagttagt 20 <210> 2167 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2167 ggttgatttt gaattaagtt 20 <210> 2168 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2168 agtttcaggt tgattttgaa 20 <210> 2169 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2169 ctggagtttc aggttgattt 20 <210> 2170 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2170 ggtgttctgg agtttcaggt 20 <210> 2171 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2171 tgggtgttct ggagtttcag 20 <210> 2172 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2172 tctgggtgtt ctggagtttc 20 <210> 2173 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2173 cttctgggtg ttctggagtt 20 <210> 2174 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2174 tacttctggg tgttctggag 20 <210> 2175 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2175 gttacttctg ggtgttctgg 20 <210> 2176 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2176 aagttacttc tgggtgttct 20 <210> 2177 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2177 gtgaagttac ttctgggtgt 20 <210> 2178 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2178 ttttaagtga agttacttct 20 <210> 2179 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2179 agttttaagt gaagttactt 20 <210> 2180 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2180 aaagttttaa gtgaagttac 20 <210> 2181 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2181 acaaaagttt taagtgaagt 20 <210> 2182 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2182 ctacaaaagt tttaagtgaa 20 <210> 2183 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2183 ttctacaaaa gttttaagtg 20 <210> 2184 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2184 tgtttttcta caaaagtttt 20 <210> 2185 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2185 cttgtttttc tacaaaagtt 20 <210> 2186 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2186 tattatcttg tttttctaca 20 <210> 2187 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2187 gatgctatta tcttgttttt 20 <210> 2188 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2188 tttgatgcta ttatcttgtt 20 <210> 2189 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2189 tctttgatgc tattatcttg 20 <210> 2190 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2190 gagaaggtct ttgatgctat 20 <210> 2191 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2191 gtctggagaa ggtctttgat 20 <210> 2192 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2192 cggtctggag aaggtctttg 20 <210> 2193 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2193 cacggtctgg agaaggtctt 20 <210> 2194 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2194 tccacggtct ggagaaggtc 20 <210> 2195 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2195 cttccacggt ctggagaagg 20 <210> 2196 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2196 gtcttccacg gtctggagaa 20 <210> 2197 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2197 tggtcttcca cggtctggag 20 <210> 2198 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2198 attggtcttc cacggtctgg 20 <210> 2199 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2199 atattggtct tccacggtct 20 <210> 2200 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2200 ttatattggt cttccacggt 20 <210> 2201 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2201 gtttatattg gtcttccacg 20 <210> 2202 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2202 ttgtttatat tggtcttcca 20 <210> 2203 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2203 aattgtttat attggtcttc 20 <210> 2204 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2204 ttaattgttt atattggtct 20 <210> 2205 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2205 gtttaattgt ttatattggt 20 <210> 2206 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2206 tggtttaatt gtttatattg 20 <210> 2207 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2207 gttggtttaa ttgtttatat 20 <210> 2208 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2208 tttgtgatcc atctattcga 20 <210> 2209 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2209 ttttgtgatc catctattcg 20 <210> 2210 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2210 attgaagttt tgtgatccat 20 <210> 2211 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2211 cattgaagtt ttgtgatcca 20 <210> 2212 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2212 tcattgaagt tttgtgatcc 20 <210> 2213 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2213 ttcattgaag ttttgtgatc 20 <210> 2214 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2214 tttcattgaa gttttgtgat 20 <210> 2215 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2215 atatttgtag ttctcccacg 20 <210> 2216 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2216 ccaaaaccat atttgtagtt 20 <210> 2217 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2217 cccaaaacca tatttgtagt 20 <210> 2218 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2218 tcccaaaacc atatttgtag 20 <210> 2219 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2219 ctcccaaaac catatttgta 20 <210> 2220 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2220 agcctcccaa aaccatattt 20 <210> 2221 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2221 caagcctccc aaaaccatat 20 <210> 2222 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2222 tcaagcctcc caaaaccata 20 <210> 2223 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2223 atcaagcctc ccaaaaccat 20 <210> 2224 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2224 catcaagcct cccaaaacca 20 <210> 2225 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2225 ccatcaagcc tcccaaaacc 20 <210> 2226 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2226 tccatcaagc ctcccaaaac 20 <210> 2227 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2227 ctccatcaag cctcccaaaa 20 <210> 2228 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2228 aaaattctcc atcaagcctc 20 <210> 2229 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2229 ccaaaattct ccatcaagcc 20 <210> 2230 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2230 accaaaattc tccatcaagc 20 <210> 2231 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2231 ccaaccaaaa ttctccatca 20 <210> 2232 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2232 cccaaccaaa attctccatc 20 <210> 2233 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2233 gcccaaccaa aattctccat 20 <210> 2234 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2234 ggcccaacca aaattctcca 20 <210> 2235 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2235 aggcccaacc aaaattctcc 20 <210> 2236 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2236 taggcccaac caaaattctc 20 <210> 2237 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2237 ctaggcccaa ccaaaattct 20 <210> 2238 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2238 tctaggccca accaaaattc 20 <210> 2239 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2239 ctctaggccc aaccaaaatt 20 <210> 2240 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2240 tctctaggcc caaccaaaat 20 <210> 2241 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2241 ttctctaggc ccaaccaaaa 20 <210> 2242 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2242 cttctctagg cccaaccaaa 20 <210> 2243 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2243 atatcttctc taggcccaac 20 <210> 2244 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2244 tatatcttct ctaggcccaa 20 <210> 2245 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2245 gtatatcttc tctaggccca 20 <210> 2246 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2246 atggagtata tcttctctag 20 <210> 2247 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2247 cactatggag tatatcttct 20 <210> 2248 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2248 tcactatgga gtatatcttc 20 <210> 2249 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2249 ttcactatgg agtatatctt 20 <210> 2250 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2250 ttgcttcact atggagtata 20 <210> 2251 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2251 attgcttcac tatggagtat 20 <210> 2252 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2252 ttagattgct tcactatgga 20 <210> 2253 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2253 attagattgc ttcactatgg 20 <210> 2254 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2254 aatttagattg cttcactatg 20 <210> 2255 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2255 taattagatt gcttcactat 20 <210> 2256 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2256 ataattagat tgcttcacta 20 <210> 2257 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2257 cataattaga ttgcttcact 20 <210> 2258 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2258 acataattag attgcttcac 20 <210> 2259 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2259 aacataatta gattgcttca 20 <210> 2260 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2260 aaacataatt agattgcttc 20 <210> 2261 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2261 aaaacataat tagattgctt 20 <210> 2262 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2262 taaaacataa ttagattgct 20 <210> 2263 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2263 gtaaaacata attagattgc 20 <210> 2264 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2264 cgtaaaacat aatttagattg 20 <210> 2265 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2265 ttccagtctt ccaactcaat 20 <210> 2266 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2266 ctttccagtc ttccaactca 20 <210> 2267 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2267 ttgttgtctt tccagtcttc 20 <210> 2268 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2268 ataatgtttg ttgtctttcc 20 <210> 2269 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2269 tataatgttt gttgtctttc 20 <210> 2270 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2270 atataatgtt tgttgtcttt 20 <210> 2271 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2271 tcaatataat gtttgttgtc 20 <210> 2272 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2272 atattcaata taatgtttgt 20 <210> 2273 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2273 aatattcaat ataatgtttg 20 <210> 2274 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2274 gaatattcaa tataatgttt 20 <210> 2275 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2275 agaatattca atataatgtt 20 <210> 2276 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2276 aagaatattc aatataatgt 20 <210> 2277 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2277 caagtaaaaa gaatattcaa 20 <210> 2278 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2278 ccaagtaaaa agaatattca 20 <210> 2279 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2279 cccaagtaaa aagaatattc 20 <210> 2280 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2280 tttcccaagt aaaaagaata 20 <210> 2281 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2281 atttcccaag taaaaagaat 20 <210> 2282 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2282 gatttcccaa gtaaaaagaa 20 <210> 2283 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2283 tgatttccca agtaaaaaga 20 <210> 2284 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2284 aatcgcaact agatgtagcg 20 <210> 2285 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2285 attctttaag gttatgtgat 20 <210> 2286 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2286 tattctttaa ggttatgtga 20 <210> 2287 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2287 acggtattct ttaaggttat 20 <210> 2288 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2288 aacggtattc tttaaggtta 20 <210> 2289 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2289 aaacggtatt ctttaaggtt 20 <210> 2290 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2290 taaacggtat tctttaaggt 20 <210> 2291 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2291 gtaaacggta ttctttaagg 20 <210> 2292 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2292 tgtaaacggt attctttaag 20 <210> 2293 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2293 atgtaaacgg tattctttaa 20 <210> 2294 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2294 aatgtaaacg gtattcttta 20 <210> 2295 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2295 aaatgtaaac ggtattcttt 20 <210> 2296 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2296 gaaatgtaaa cggtattctt 20 <210> 2297 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2297 agaaatgtaa acggtattct 20 <210> 2298 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2298 gagaaatgta aacggtattc 20 <210> 2299 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2299 tgagaaatgt aaacggtatt 20 <210> 2300 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2300 ttgagaaatg taaacggtat 20 <210> 2301 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2301 attgagaaat gtaaacggta 20 <210> 2302 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2302 gattgagaaa tgtaaacggt 20 <210> 2303 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2303 tgattgagaa atgtaaacgg 20 <210> 2304 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2304 ttgattgaga aatgtaaacg 20 <210> 2305 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2305 tttgattgag aaatgtaaac 20 <210> 2306 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2306 ttttgattga gaaatgtaaa 20 <210> 2307 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2307 attttgattg agaaatgtaa 20 <210> 2308 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2308 aattttgatt gagaaatgta 20 <210> 2309 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2309 gaattttgat tgagaaatgt 20 <210> 2310 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2310 agaattttga ttgagaaatg 20 <210> 2311 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2311 aagaattttg attgagaaat 20 <210> 2312 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2312 ataagaattt tgattgagaa 20 <210> 2313 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2313 tataagaatt ttgattgaga 20 <210> 2314 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2314 ttataagaat tttgattgag 20 <210> 2315 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2315 attataagaa ttttgattga 20 <210> 2316 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2316 tattataaga attttgattg 20 <210> 2317 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2317 gtattataag aattttgatt 20 <210> 2318 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2318 agtattataa gaattttgat 20 <210> 2319 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2319 tagtattata agaattttga 20 <210> 2320 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2320 aaaacaaata gtattataag 20 <210> 2321 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2321 taaaacaaat agtattataa 20 <210> 2322 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2322 attcccacat cacaaaattt 20 <210> 2323 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2323 gattcccaca tcacaaaatt 20 <210> 2324 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2324 tgattcccac atcacaaaat 20 <210> 2325 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2325 aaattgattc ccacatcaca 20 <210> 2326 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2326 aaaattgatt cccacatcac 20 <210> 2327 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2327 atctaaaatt gattcccaca 20 <210> 2328 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2328 gaccatctaa aattgattcc 20 <210> 2329 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2329 tgaccatcta aaattgattc 20 <210> 2330 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2330 gtgaccatct aaaattgatt 20 <210> 2331 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2331 tgtgaccatc taaaattgat 20 <210> 2332 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2332 ttgtgaccat ctaaaattga 20 <210> 2333 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2333 attgtgacca tctaaaattg 20 <210> 2334 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2334 gattgtgacc atctaaaatt 20 <210> 2335 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2335 agattgtgac catctaaaat 20 <210> 2336 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2336 tagattgtga ccatctaaaa 20 <210> 2337 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2337 ctagattgtg accatctaaa 20 <210> 2338 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2338 tctagattgt gaccatctaa 20 <210> 2339 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2339 atctagattg tgaccattcta 20 <210> 2340 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2340 aatctagatt gtgaccatct 20 <210> 2341 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2341 taatctagat tgtgaccatc 20 <210> 2342 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2342 ataatctaga ttgtgaccat 20 <210> 2343 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2343 tataatctag attgtgacca 20 <210> 2344 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2344 ttataatcta gattgtgacc 20 <210> 2345 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2345 tgattataat ctagattgtg 20 <210> 2346 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2346 ttgattataa tctagattgt 20 <210> 2347 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2347 attgattata atctagattg 20 <210> 2348 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2348 tattgattat aatctagatt 20 <210> 2349 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2349 ctattgatta taatctagat 20 <210> 2350 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2350 cctattgatt ataatctaga 20 <210> 2351 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2351 acctattgat tataatctag 20 <210> 2352 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2352 cacctattga ttataatcta 20 <210> 2353 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2353 tcacctattg attataatct 20 <210> 2354 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2354 ttcacctatt gattataatc 20 <210> 2355 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2355 gttcacctat tgattataat 20 <210> 2356 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2356 agttcaccta ttgattataa 20 <210> 2357 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2357 aagttcacct attgattata 20 <210> 2358 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2358 ataagttcac ctattgatta 20 <210> 2359 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2359 aataagttca cctattgatt 20 <210> 2360 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2360 taataagttc acctattgat 20 <210> 2361 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2361 ttaataagtt cacctattga 20 <210> 2362 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2362 tgagtattct cgacttacct 20 <210> 2363 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2363 aagtgagtat tctcgactta 20 <210> 2364 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2364 attaagtgag tattctcgac 20 <210> 2365 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2365 ccagaattaa gtgagtattc 20 <210> 2366 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2366 tgggttgctt tcttaccaga 20 <210> 2367 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2367 aaatgggttg ctttcttacc 20 <210> 2368 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2368 tacaaatggg ttgctttctt 20 <210> 2369 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2369 aagtacaaat gggttgcttt 20 <210> 2370 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2370 gtaaatacaa gtacaaatgg 20 <210> 2371 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2371 ttgctggtaa atacaagtac 20 <210> 2372 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2372 taaggattgc tggtaaatac 20 <210> 2373 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2373 ttttaaggat tgctggtaaa 20 <210> 2374 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2374 gaagcttcat tttaaggatt 20 <210> 2375 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2375 taggaagctt cattttaagg 20 <210> 2376 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2376 tagtaggaag cttcatttta 20 <210> 2377 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2377 ttgagttagt aggaagcttc 20 <210> 2378 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2378 attgctattg agttagtagg 20 <210> 2379 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2379 cttattgcta ttgagttagt 20 <210> 2380 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2380 attgtctttat tgctattgag 20 <210> 2381 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2381 actattgtct tattgctatt 20 <210> 2382 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2382 ttcactattg tcttattgct 20 <210> 2383 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2383 acattcacta ttgtcttatt 20 <210> 2384 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2384 taaacattca ctattgtctt 20 <210> 2385 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2385 cattaaacat tcactattgt 20 <210> 2386 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2386 gttttcatta aacattcact 20 <210> 2387 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2387 aaatactgtt ttcattaaac 20 <210> 2388 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2388 aaagtattta taaaatactg 20 <210> 2389 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2389 cctttttatt aaagttatta 20 <210> 2390 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2390 caatcctttt tattaaagta 20 <210> 2391 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2391 cttcatcaca atccttttta 20 <210> 2392 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2392 gttcttcatc acaatccttt 20 <210> 2393 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2393 attgttcttc atcacaatcc 20 <210> 2394 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2394 tagattgttc ttcatcacaa 20 <210> 2395 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2395 aaatagattg ttcttcatca 20 <210> 2396 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2396 aacaaatata aatagattgt 20 <210> 2397 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2397 caaataacaa atataaatag 20 <210> 2398 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2398 tggaattaaa aacaaataac 20 <210> 2399 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2399 ttattggaat taaaaacaaa 20 <210> 2400 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2400 tttttattgg aattaaaaac 20 <210> 2401 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2401 taataacttt tttctgtaat 20 <210> 2402 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2402 gttcttaata acttttttct 20 <210> 2403 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2403 aaaagcatgg ttcttaataa 20 <210> 2404 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2404 aaatttaaaa gcatggttct 20 <210> 2405 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2405 aggaataaat ttaaaaaatc 20 <210> 2406 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2406 agacaggaat aaatttaaaa 20 <210> 2407 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2407 aaaagacagg aataaattta 20 <210> 2408 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2408 ctttctttgt agaaaaagac 20 <210> 2409 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2409 atgctttctt tgtagaaaaa 20 <210> 2410 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2410 tttgcttaat gtatgctttc 20 <210> 2411 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2411 gtatttgctt aatgtatgct 20 <210> 2412 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2412 ttggtatttg cttaatgtat 20 <210> 2413 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2413 cctttggtat ttgcttaatg 20 <210> 2414 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2414 tggcctttgg tatttgctta 20 <210> 2415 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2415 taaacctggc ctttggtatt 20 <210> 2416 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2416 atgtaaacct ggcctttggt 20 <210> 2417 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2417 caaatgtaaa cctggccttt 20 <210> 2418 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2418 cttcaaatgt aaacctggcc 20 <210> 2419 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2419 tttcttcaaa tgtaaacctg 20 <210> 2420 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2420 tgtcactttc ttcaaatgta 20 <210> 2421 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2421 taatgtcact ttcttcaaat 20 <210> 2422 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2422 aataataatg tcactttctt 20 <210> 2423 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2423 gagtaataat aatgtcactt 20 <210> 2424 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2424 gacttgagta ataataatgt 20 <210> 2425 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2425 cctagagact tgagtaataa 20 <210> 2426 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2426 attcctagag acttgagtaa 20 <210> 2427 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2427 aagtattcct agagacttga 20 <210> 2428 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2428 tgtgttaagt attcctagag 20 <210> 2429 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2429 agagatgtgt taagtattcc 20 <210> 2430 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2430 gtcaagagat gtgttaagta 20 <210> 2431 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2431 acagtcaaga gatgtgttaa 20 <210> 2432 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2432 tatacagtca agagatgtgt 20 <210> 2433 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2433 ccatatacag tcaagagatg 20 <210> 2434 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2434 gtaagttgaa ctaactactg 20 <210> 2435 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2435 tgagtaagtt gaactaacta 20 <210> 2436 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2436 taatgagtaa gttgaactaa 20 <210> 2437 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2437 aggttaatct tcctaatacg 20 <210> 2438 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2438 ataaccaggt taatcttcct 20 <210> 2439 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2439 atgataacca ggttaatctt 20 <210> 2440 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2440 aacaatgata accaggttaa 20 <210> 2441 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2441 taaaacaatg ataaccaggt 20 <210> 2442 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2442 gtataaaaca atgataacca 20 <210> 2443 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2443 cgaatactca tatatatttc 20 <210> 2444 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2444 atacgaatac tcatatatat 20 <210> 2445 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2445 tttatacgaa tactcatata 20 <210> 2446 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2446 atatttatac gaatactcat 20 <210> 2447 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2447 gtattatatt tatacgaata 20 <210> 2448 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2448 aaaagtatta tattatatacg 20 <210> 2449 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2449 ggtaaaagta ttatatttat 20 <210> 2450 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2450 acaaggtaaa agtattatat 20 <210> 2451 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2451 taaacaaggt aaaagtatta 20 <210> 2452 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2452 acataaacaa ggtaaaagta 20 <210> 2453 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2453 ttgagtaaat acataaacaa 20 <210> 2454 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2454 gagaatattg agtaaataca 20 <210> 2455 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2455 aaggagaata ttgagtaaat 20 <210> 2456 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2456 gaaaaggaga atattgagta 20 <210> 2457 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2457 gaggaaaagg agaatattga 20 <210> 2458 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2458 ttagaggaaa aggagaatat 20 <210> 2459 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2459 attattttag aggaaaagga 20 <210> 2460 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2460 cagattattt tagaggaaaa 20 <210> 2461 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2461 cttcagatta ttttagagga 20 <210> 2462 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2462 tagtcacttc agattatttt 20 <210> 2463 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2463 taatagtcac ttcagattat 20 <210> 2464 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2464 tgataatagt cacttcagat 20 <210> 2465 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2465 tattgataat agtcacttca 20 <210> 2466 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2466 acttattgat aatagtcact 20 <210> 2467 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2467 taaacttatt gataatagtc 20 <210> 2468 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2468 tagtaaactt attgataata 20 <210> 2469 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2469 gcatagtaaa cttattgata 20 <210> 2470 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2470 ttggcatagt aaacttattg 20 <210> 2471 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2471 attttggcat agtaaactta 20 <210> 2472 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2472 tgaattttgg catagtaaac 20 <210> 2473 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2473 ttaatgaatt ttggcatagt 20 <210> 2474 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2474 caattaatga attttggcat 20 <210> 2475 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2475 aaaggcaatt aatgaatttt 20 <210> 2476 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2476 gtgaaaggca attaatgaat 20 <210> 2477 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2477 ttaagtgaaa ggcaattaat 20 <210> 2478 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2478 aagttaagtg aaaggcaatt 20 <210> 2479 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2479 ccaaaagtta agtgaaaggc 20 <210> 2480 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2480 gtcccaaaag ttaagtgaaa 20 <210> 2481 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2481 atggtcccaa aagttaagtg 20 <210> 2482 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2482 attatggtcc caaaagttaa 20 <210> 2483 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2483 tttattatgg tcccaaaagt 20 <210> 2484 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2484 ttattattta ttatggtccc 20 <210> 2485 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2485 atggcaatac attttattat 20 <210> 2486 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2486 gttatggcaa tacattttat 20 <210> 2487 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2487 taatgttatg gcaatacatt 20 <210> 2488 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2488 tattaatgtt atggcaatac 20 <210> 2489 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2489 gtttattaat gttatggcaa 20 <210> 2490 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2490 gtagtttatt aatgttatgg 20 <210> 2491 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2491 aaggtagttt attaatgtta 20 <210> 2492 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2492 tgtaaggtag tttattaatg 20 <210> 2493 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2493 ttttgtaagg tagtttatta 20 <210> 2494 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2494 tggttttgta aggtagttta 20 <210> 2495 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2495 tggtggtttt gtaaggtagt 20 <210> 2496 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2496 aattggtggt tttgtaaggt 20 <210> 2497 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2497 tttaattggt ggttttgtaa 20 <210> 2498 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2498 ttgattttaa ttggtggttt 20 <210> 2499 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2499 tgtttgattt taattggtgg 20 <210> 2500 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2500 atgtaaataa cacttttttg 20 <210> 2501 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2501 cagatgtaaa taacactttt 20 <210> 2502 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2502 tgacagatgt aaataacact 20 <210> 2503 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2503 atgttgacag atgtaaataa 20 <210> 2504 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2504 tttatgttga cagatgtaaa 20 <210> 2505 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2505 agatttatgt tgacagatgt 20 <210> 2506 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2506 agtagattta tgttgacaga 20 <210> 2507 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2507 tttagtagat ttatgttgac 20 <210> 2508 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2508 atttttagta gattatgtt 20 <210> 2509 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2509 catgtatttt tagtagattt 20 <210> 2510 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2510 gaaatcatgt atttttagta 20 <210> 2511 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2511 attgtatttg atggatatct 20 <210> 2512 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2512 gatacattgt atttgatgga 20 <210> 2513 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2513 taggttgata cattgtattt 20 <210> 2514 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2514 cagtttaggt tgatacattg 20 <210> 2515 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2515 gcatccagtt taggttgata 20 <210> 2516 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2516 ccccagcatc cagtttaggt 20 <210> 2517 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2517 aagaacccca gcatccagtt 20 <210> 2518 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2518 gtgtaaaaag aaccccagca 20 <210> 2519 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2519 atagggtgta aaaagaaccc 20 <210> 2520 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2520 cttttatagg gtgtaaaaag 20 <210> 2521 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2521 tatgtctttt atagggtgta 20 <210> 2522 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2522 ttaggtatgt cttttatagg 20 <210> 2523 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2523 ttgtcttagg tatgtctttt 20 <210> 2524 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2524 ctctgattgt cttaggtatg 20 <210> 2525 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2525 tatttctctg attgtcttag 20 <210> 2526 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2526 tcaagtccat atttgtattt 20 <210> 2527 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2527 aataatcaag tccatatttg 20 <210> 2528 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2528 ttatctaata atcaagtcca 20 <210> 2529 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2529 ctatattatc taataatcaa 20 <210> 2530 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2530 taaaccttct atattatcta 20 <210> 2531 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2531 aattaataaa ccttctatat 20 <210> 2532 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2532 taagtacagg ttggacactg 20 <210> 2533 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2533 gttattaagt acaggttgga 20 <210> 2534 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2534 tgtgagttat taagtacagg 20 <210> 2535 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2535 aaatctgtga gttattaagt 20 <210> 2536 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2536 gttttaaaaa tctgtgagtt 20 <210> 2537 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2537 caaaattctc ctgaaaagaa 20 <210> 2538 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2538 cccaaccaaa attctcctga 20 <210> 2539 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2539 acctgaataa ccctctggac 20 <210> 2540 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2540 aagatacctg aataaccctc 20 <210> 2541 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2541 agaaaaagat acctgaataa 20 <210> 2542 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2542 tggtatcaga aaaagatacc 20 <210> 2543 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2543 agtattggta tcagaaaaag 20 <210> 2544 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2544 aataaagtat tggtatcaga 20 <210> 2545 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2545 atgaaaataa agtattggta 20 <210> 2546 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2546 agatactttg aagatatgaa 20 <210> 2547 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2547 tgggaagata ctttgaagat 20 <210> 2548 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2548 ctaataatgt gggaagatac 20 <210> 2549 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2549 cattgcagat aatagctaat 20 <210> 2550 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2550 aagttgtcat tgcagataat 20 <210> 2551 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2551 ttttaaaagt tgtcattgca 20 <210> 2552 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2552 attcggattt ttaaaagttg 20 <210> 2553 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2553 ttatttggga ttcggatttt 20 <210> 2554 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2554 ttatagttaa gaggttttcg 20 <210> 2555 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2555 tttcattata gttaagaggt 20 <210> 2556 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2556 gaacactttc attatagtta 20 <210> 2557 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2557 gaactagaat gaacactttc 20 <210> 2558 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2558 tgattgaact agaatgaaca 20 <210> 2559 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2559 atacctgatt gaactagaat 20 <210> 2560 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2560 gtaaaatacc tgattgaact 20 <210> 2561 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2561 tagaggtaaa atacctgatt 20 <210> 2562 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2562 aagattagag gtaaaatacc 20 <210> 2563 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2563 tgaggaagat tagaggtaaa 20 <210> 2564 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2564 gaaaatctga ggaagattag 20 <210> 2565 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2565 aaatagaaaa tctgaggaag 20 <210> 2566 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2566 atctatacac taccaaaaaa 20 <210> 2567 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2567 aaataatcta tacactacca 20 <210> 2568 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2568 aaataatctg tataaataat 20 <210> 2569 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2569 cccaatttta aataatctgt 20 <210> 2570 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2570 taagtcccaa ttttaaataa 20 <210> 2571 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2571 tctgtataag tcccaatttt 20 <210> 2572 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2572 aataatctgt ataagtccca 20 <210> 2573 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2573 agttttaaat aatctgtata 20 <210> 2574 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2574 atcccagttt taaataatct 20 <210> 2575 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2575 catgtatccc agttttaaat 20 <210> 2576 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2576 tagatgcatg tatcccagtt 20 <210> 2577 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2577 tgttttagat gcatgtatcc 20 <210> 2578 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2578 tacagtgttt tagatgcatg 20 <210> 2579 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2579 aatattacag tgttttagat 20 <210> 2580 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2580 cttataaata ttacagtgtt 20 <210> 2581 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2581 cttcctttct tataaatatt 20 <210> 2582 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2582 tttatcttcc tttcttataa 20 <210> 2583 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2583 ttccccgtaa gtttatcttc 20 <210> 2584 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2584 tgtatttccc cgtaagttta 20 <210> 2585 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2585 gttactgtat ttccccgtaa 20 <210> 2586 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2586 tgttttcttc tggaagcaga 20 <210> 2587 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2587 cagacctaga cttcttaact 20 <210> 2588 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2588 agcagaccta gacttcttaa 20 <210> 2589 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2589 ttttcttctg gaagcagacc 20 <210> 2590 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2590 aaacatatat acagcttgt 20 <210> 2591 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2591 ttaaacatat atacatgctt 20 <210> 2592 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2592 gtttattgaa ttttaaacat 20 <210> 2593 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2593 ttgtttattg aattttaaac 20 <210> 2594 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2594 ctttgtttat tgaattttaa 20 <210> 2595 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2595 gtctttgttt attgaatttt 20 <210> 2596 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2596 gggtctttgt ttattgaatt 20 <210> 2597 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2597 ctgggtcttt gtttattgaa 20 <210> 2598 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2598 gactgggtct ttgtttattg 20 <210> 2599 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2599 tttctataat ttagggactg 20 <210> 2600 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2600 aatttctata atttagggac 20 <210> 2601 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2601 taaatttcta taatttaggg 20 <210> 2602 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2602 caagaataat ttaaatttct 20 <210> 2603 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2603 gataaacatg caagaataat 20 <210> 2604 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2604 tcgataaaca tgcaagaata 20 <210> 2605 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2605 tgtcgataaa catgcaagaa 20 <210> 2606 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2606 gatgtcgata aacatgcaag 20 <210> 2607 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2607 gtgatgtcga taaacatgca 20 <210> 2608 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2608 ttgtgatgtc gataaacatg 20 <210> 2609 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2609 tgttgtgatg tcgataaaca 20 <210> 2610 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2610 tctgttgtga tgtcgataaa 20 <210> 2611 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2611 gatctgttgt gatgtcgata 20 <210> 2612 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2612 gggatctgtt gtgatgtcga 20 <210> 2613 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2613 tagggatctg ttgtgatgtc 20 <210> 2614 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2614 tttagggatc tgttgtgatg 20 <210> 2615 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2615 gattaggga tctgttgtga 20 <210> 2616 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2616 atctaatctt tagggattta 20 <210> 2617 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2617 tttgtatcta atctttaggg 20 <210> 2618 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2618 aatttgtatc taatctttag 20 <210> 2619 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2619 gtggtaaaaa atttgtatct 20 <210> 2620 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2620 ctgtggtaaa aaatttgtat 20 <210> 2621 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2621 tactgtggta aaaaatttgt 20 <210> 2622 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2622 gatactgtgg taaaaaattt 20 <210> 2623 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2623 agtgatactg tggtaaaaaa 20 <210> 2624 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2624 caagtgatac tgtggtaaaa 20 <210> 2625 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2625 gacaagtgat actgtggtaa 20 <210> 2626 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2626 ctgacaagtg atactgtggt 20 <210> 2627 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2627 ttctgacaag tgatactgtg 20 <210> 2628 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2628 aattctgaca agtgatactg 20 <210> 2629 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2629 ataaattctg acaagtgata 20 <210> 2630 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2630 ctggcagttt taaaaaatca 20 <210> 2631 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2631 ttcttactgg cagttttaaa 20 <210> 2632 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2632 atttcttact ggcagtttta 20 <210> 2633 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2633 aaatttctta ctggcagttt 20 <210> 2634 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2634 tttaaaattt cttactggca 20 <210> 2635 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2635 ttaatttaaa atttcttact 20 <210> 2636 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2636 caaatgggtt taatttaaaa 20 <210> 2637 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2637 aacaaatggg tttaatttaa 20 <210> 2638 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2638 ttaacaaatg ggtttaattt 20 <210> 2639 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2639 ctttaacaaa tgggtttaat 20 <210> 2640 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2640 tcctttaaca aatgggttta 20 <210> 2641 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2641 ctatatcctt taacaaatgg 20 <210> 2642 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2642 gggcactata tcctttaaca 20 <210> 2643 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2643 ttgggcacta tatcctttaa 20 <210> 2644 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2644 tataacttgg gcactatatc 20 <210> 2645 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2645 catataactt gggcactata 20 <210> 2646 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2646 tcaccatata acttgggcac 20 <210> 2647 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2647 taggtcacca tataacttgg 20 <210> 2648 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2648 ggtaggtcac catataactt 20 <210> 2649 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2649 aaggtaggtc accatataac 20 <210> 2650 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2650 caaaggtagg tcaccatata 20 <210> 2651 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2651 gtattgacaa aggtaggtca 20 <210> 2652 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2652 aagtattgac aaaggtaggt 20 <210> 2653 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2653 ctaagtattg acaaaggtag 20 <210> 2654 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2654 tgctaagtat tgacaaaggt 20 <210> 2655 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2655 aatgctaagt attgacaaag 20 <210> 2656 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2656 cataatgcta agtattgaca 20 <210> 2657 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2657 tacataatgc taagtattga 20 <210> 2658 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2658 aatacataat gctaagtatt 20 <210> 2659 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2659 gaaatacata atgctaagta 20 <210> 2660 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2660 tttgaaatac ataatgctaa 20 <210> 2661 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2661 ggataatttg aaatacataa 20 <210> 2662 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2662 ttggataatt tgaaatacat 20 <210> 2663 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2663 tattggataa tttgaaatac 20 <210> 2664 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2664 atccagttaa agcttgtaaa 20 <210> 2665 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2665 tcatgatcca gttaaagctt 20 <210> 2666 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2666 tttactcatg atccagttaa 20 <210> 2667 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2667 gataatttta ctcatgatcc 20 <210> 2668 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2668 gatgtgataa ttttactcat 20 <210> 2669 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2669 atgctgatgt gataatttta 20 <210> 2670 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2670 cagttatgct gatgtgataa 20 <210> 2671 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2671 tttaacagtt atgctgatgt 20 <210> 2672 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2672 gcaattttaa cagttatgct 20 <210> 2673 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2673 agagcctgca attttaacag 20 <210> 2674 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2674 gcttcagagc ctgcaatttt 20 <210> 2675 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2675 tattagcttc agagcctgca 20 <210> 2676 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2676 tagtttatta gcttcagagc 20 <210> 2677 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2677 gcaggtagtt tattagcttc 20 <210> 2678 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2678 taaatgcagg tagtttatta 20 <210> 2679 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2679 atggtttaaa tgcaggtagt 20 <210> 2680 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2680 gagccatggt ttaaatgcag 20 <210> 2681 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2681 ttttagagcc atggtttaaa 20 <210> 2682 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2682 caaagtttta gagccatggt 20 <210> 2683 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2683 tcacacaaag ttttagagcc 20 <210> 2684 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2684 caaggtcaca caaagtttta 20 <210> 2685 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2685 gggtgaagta atttattcaa 20 <210> 2686 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2686 gtgaggaaac tgagagataa 20 <210> 2687 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2687 tgtagtatat gtgaggaaac 20 <210> 2688 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2688 atctttgtag tatatgtgag 20 <210> 2689 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2689 ttattatctt tgtagtatat 20 <210> 2690 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2690 ttctgttatt atctttgtag 20 <210> 2691 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2691 ataagttctg ttattatctt 20 <210> 2692 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2692 atcctataag ttctgttatt 20 <210> 2693 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2693 caataatcct ataagttctg 20 <210> 2694 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2694 taagatgaca ttggctgcta 20 <210> 2695 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2695 tttagtaaga tgacattggc 20 <210> 2696 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2696 ttgaatttta gtaagatgac 20 <210> 2697 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2697 ctaatttgaa ttttagtaag 20 <210> 2698 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2698 catgatctaa tttgaatttt 20 <210> 2699 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2699 caaagagaaa catgatctaa 20 <210> 2700 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2700 gttttgagca aagagaaaca 20 <210> 2701 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2701 gtgtggtttt gagcaaagag 20 <210> 2702 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2702 agctattgtg tggttttgag 20 <210> 2703 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2703 tgaaatggaa agctattgtg 20 <210> 2704 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2704 tatgagtgaa atggaaagct 20 <210> 2705 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2705 aaagagccaa tatgagtgaa 20 <210> 2706 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2706 ttggtctaaa gagccaatat 20 <210> 2707 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2707 ggtaatcttg gtctaaagag 20 <210> 2708 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2708 gtgagatgac gaagggttgg 20 <210> 2709 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2709 agtcagtgag atgacgaagg 20 <210> 2710 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2710 ggtgaagtca gtgagatgac 20 <210> 2711 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2711 gtagaggagg tgaagtcagt 20 <210> 2712 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2712 aactagagta gaggaggtga 20 <210> 2713 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2713 agaataacta gagtagagga 20 <210> 2714 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2714 cggtcagaat aactagagta 20 <210> 2715 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2715 taaagcggtc agaataacta 20 <210> 2716 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2716 actggtaaag cggtcagaat 20 <210> 2717 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2717 tgaatactgg taaagcggtc 20 <210> 2718 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2718 tgtgtttgaa tactggtaaa 20 <210> 2719 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2719 agtatgtttg atgtgtttga 20 <210> 2720 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2720 gtggcagtat gtttgatgtg 20 <210> 2721 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2721 ttgaggtggc agtatgtttg 20 <210> 2722 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2722 aggctttgag gtggcagtat 20 <210> 2723 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2723 ggcaaaggct ttgaggtggc 20 <210> 2724 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2724 aacaagggca aaggctttga 20 <210> 2725 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2725 tagaggaaac aacaagggca 20 <210> 2726 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2726 ccagttagag gaaacaacaa 20 <210> 2727 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2727 gataccaggg cagaagagcg 20 <210> 2728 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2728 aaatcagaga gtgggccacg 20 <210> 2729 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2729 cctaagggaa atcagagagt 20 <210> 2730 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2730 acgaccctaa gggaaatcag 20 <210> 2731 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2731 tgataacgac cctaagggaa 20 <210> 2732 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2732 ttttgtttga taacgaccct 20 <210> 2733 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2733 gtcttcattg ggaatttttt 20 <210> 2734 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2734 tgtaagtctt cattgggaat 20 <210> 2735 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2735 gaccttgtaa gtcttcattg 20 <210> 2736 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2736 taagtgacct tgtaagtctt 20 <210> 2737 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2737 ttggttaagt gaccttgtaa 20 <210> 2738 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2738 tgatttttgg ttaagtgacc 20 <210> 2739 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2739 ggttgtgatt tttggttaag 20 <210> 2740 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2740 caggcggttg tgatttttgg 20 <210> 2741 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2741 gggaccaggc ggttgtgatt 20 <210> 2742 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2742 ctaaggaagt agaagttttc 20 <210> 2743 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2743 agtagctaag gaagtagaag 20 <210> 2744 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2744 caggagaaaa gtagctaagg 20 <210> 2745 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2745 gtgtgcagga gaaaagtagc 20 <210> 2746 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2746 taaaggtgag tgtgcaggag 20 <210> 2747 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2747 atgttaaata aaggtgagtg 20 <210> 2748 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2748 atgttatgtt aaataaaggt 20 <210> 2749 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2749 aatttatgtt atgttaaata 20 <210> 2750 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2750 taactaaaat ttatgttatg 20 <210> 2751 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2751 gataaataac taaaatttat 20 <210> 2752 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2752 tttagtgcag gaatagaaga 20 <210> 2753 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2753 aatccctgta ttcacagagc 20 <210> 2754 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2754 gaaaaaatcc ctgtattcac 20 <210> 2755 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2755 taatggaaaa aatccctgta 20 <210> 2756 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2756 aaatatgaag ataatggaaa 20 <210> 2757 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2757 ataatggaaa atatgaagat 20 <210> 2758 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2758 tatacaaata atggaaaata 20 <210> 2759 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2759 ttctggagta tatacaaata 20 <210> 2760 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2760 attctatatt ctggagtata 20 <210> 2761 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2761 ccatacagta ttctatattc 20 <210> 2762 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2762 ctgtgtgcca tacagtattc 20 <210> 2763 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2763 gcctactgtg tgccatacag 20 <210> 2764 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2764 agaaatgcct actgtgtgcc 20 <210> 2765 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2765 tcaacagaaa tgcctactgt 20 <210> 2766 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2766 attaattcaa cagaaatgcc 20 <210> 2767 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2767 gacattacat ttattaattc 20 <210> 2768 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2768 gtgaatatga cattacattt 20 <210> 2769 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2769 cttctgtgtg aatatgacat 20 <210> 2770 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2770 acacgcttct gtgtgaatat 20 <210> 2771 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2771 atagcacacg cttctgtgtg 20 <210> 2772 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2772 taatcatagc acacgcttct 20 <210> 2773 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2773 aataataatc atagcacacg 20 <210> 2774 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2774 ccaagtaata ataatcatag 20 <210> 2775 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2775 ctagtaatcc aagtaataat 20 <210> 2776 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2776 tatttctagt aatccaagta 20 <210> 2777 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2777 cacactattt ctagtaatcc 20 <210> 2778 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2778 ttatgaggca cactatttct 20 <210> 2779 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2779 tttaattatg aggcacacta 20 <210> 2780 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2780 gttgaccttt aattatgagg 20 <210> 2781 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2781 ttacattgtt gaatgttgac 20 <210> 2782 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2782 attaattaca ttgttgaatg 20 <210> 2783 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2783 tgtagattaa ttacattgtt 20 <210> 2784 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2784 tacattgtag attaattaca 20 <210> 2785 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2785 agatgtttac attgtagatt 20 <210> 2786 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2786 ttcaccagat gtttacattg 20 <210> 2787 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2787 gtcacttcac cagatgttta 20 <210> 2788 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2788 cctctgtcac ttcaccagat 20 <210> 2789 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2789 gcttccctct gtcacttcac 20 <210> 2790 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2790 caagtgcttc cctctgtcac 20 <210> 2791 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2791 gcttttttct aaacaagtgc 20 <210> 2792 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2792 acatagcttt tttctaaaca 20 <210> 2793 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2793 ttctgacata gcttttttct 20 <210> 2794 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2794 atggattctg acatagcttt 20 <210> 2795 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2795 aatacatgga ttctgacata 20 <210> 2796 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2796 attagaatac atggattctg 20 <210> 2797 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2797 ttgtactgca tattagaata 20 <210> 2798 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2798 aactattgta ctgcatatta 20 <210> 2799 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2799 ttttaaacta ttgtactgca 20 <210> 2800 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2800 tgagagtatt attaatattt 20 <210> 2801 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2801 gctgtttgag agtattatta 20 <210> 2802 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2802 gaatagctgt ttgagagtat 20 <210> 2803 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2803 cctcttgaat agctgtttga 20 <210> 2804 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2804 tgaatcctct tgaatagctg 20 <210> 2805 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2805 ttttttgaat cctcttgaat 20 <210> 2806 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2806 ttatgttttt tgaatcctct 20 <210> 2807 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2807 gtttatatta tgttttttga 20 <210> 2808 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2808 tctgagttta tattatgttt 20 <210> 2809 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2809 cagtttctct gagtttatat 20 <210> 2810 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2810 gtttaccagt ttctctgagt 20 <210> 2811 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2811 attttgttta ccagtttctc 20 <210> 2812 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2812 aaatgatttt gtttaccagt 20 <210> 2813 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2813 ctcttgaaaa tgattttgtt 20 <210> 2814 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2814 tatatctctt gaaaatgatt 20 <210> 2815 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2815 caggttggca agtttgtttg 20 <210> 2816 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2816 gttggcaggt tggcaagttt 20 <210> 2817 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2817 atatctgtag atgttggcag 20 <210> 2818 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2818 taaacatatc tgtagatgtt 20 <210> 2819 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2819 acctgtaaac atatctgtag 20 <210> 2820 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2820 ttttgacctg taaacatatc 20 <210> 2821 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2821 ataatttttg acctgtaaac 20 <210> 2822 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2822 taatttgata atttttgacc 20 <210> 2823 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2823 ttcttgataa tttgataatt 20 <210> 2824 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2824 accaggcttt cttgataatt 20 <210> 2825 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2825 tttgaaccag gctttcttga 20 <210> 2826 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2826 agacataata cataatttga 20 <210> 2827 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2827 ctgtgataaa gacataatac 20 <210> 2828 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2828 cagacctgtg ataaagacat 20 <210> 2829 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2829 atcttcagac ctgtgataaa 20 <210> 2830 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2830 tactgatctt cagacctgtg 20 <210> 2831 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2831 ttaataattt tcagttttag 20 <210> 2832 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2832 taagtttaat aattttcagt 20 <210> 2833 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2833 ttcagatttt aagtttaata 20 <210> 2834 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2834 tatatttgat attctgttca 20 <210> 2835 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2835 atattgtaat gtattctttt 20 <210> 2836 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2836 ttagaatatt gtaatgtatt 20 <210> 2837 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2837 tttgcttaga atattgtaat 20 <210> 2838 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2838 actgctttgc ttagaatatt 20 <210> 2839 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2839 aagtagagac tgctttgctt 20 <210> 2840 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2840 gcaaggccaa aagtagagac 20 <210> 2841 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2841 acagagcaag gccaaaagta 20 <210> 2842 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2842 ggaaaacaga gcaaggccaa 20 <210> 2843 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2843 tggtcggaaa acagagcaag 20 <210> 2844 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2844 gacattggtc ggaaaacaga 20 <210> 2845 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2845 aagcagacat tggtcggaaa 20 <210> 2846 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2846 caaggcaaaa aagcagacat 20 <210> 2847 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2847 ataaagcaag gcaaaaaagc 20 <210> 2848 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2848 cattattaa taagataaaa 20 <210> 2849 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2849 aaatatttaa tcagggacat 20 <210> 2850 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2850 tgttctcaaa atatttaatc 20 <210> 2851 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2851 gattacctgt tctcaaaata 20 <210> 2852 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2852 gattgtacag attacctgtt 20 <210> 2853 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2853 attcagattg tacagatac 20 <210> 2854 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2854 aaacagtgtt attcagattg 20 <210> 2855 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2855 tagataaaca gtgttattca 20 <210> 2856 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2856 atatttagat aaacagtgtt 20 <210> 2857 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2857 gtttgatatt tagataaaca 20 <210> 2858 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2858 aacggtgttt gatatttaga 20 <210> 2859 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2859 gttataacgg tgtttgatat 20 <210> 2860 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2860 ataatgttat aacggtgttt 20 <210> 2861 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2861 agttcataat gttataacgg 20 <210> 2862 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2862 ctttcagttc ataatgttat 20 <210> 2863 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2863 agtacagttt gtctttcagt 20 <210> 2864 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2864 tcagaagtac agtttgtctt 20 <210> 2865 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2865 ggatgtcaga agtacagttt 20 <210> 2866 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2866 gagtaaggat gtcagaagta 20 <210> 2867 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2867 gaaatctgag taaggatgtc 20 <210> 2868 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2868 tactgaatat acaattaggg 20 <210> 2869 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2869 aatgatactg aatatacaat 20 <210> 2870 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2870 gaatataaat ctgtttttta 20 <210> 2871 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2871 taaaagaata taaatctgtt 20 <210> 2872 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2872 gctgataaaa gaatataaat 20 <210> 2873 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2873 ccttctgagc tgataaaaga 20 <210> 2874 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2874 ctagtccttc tgagctgata 20 <210> 2875 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2875 ttaccatcat gttttacatt 20 <210> 2876 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2876 caaagtgtct taccatcatg 20 <210> 2877 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2877 aaacccacca aagtgtctta 20 <210> 2878 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2878 agaaggaaac ccaccaaagt 20 <210> 2879 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2879 aataatagct tcaagaagga 20 <210> 2880 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2880 aatttgataa taatagcttc 20 <210> 2881 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2881 tagggaattt gataataata 20 <210> 2882 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2882 aagaataggg aatttgataa 20 <210> 2883 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2883 gtcctaagaa tagggaattt 20 <210> 2884 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2884 tagaacaagt cctaagaata 20 <210> 2885 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2885 ttagtctaga acaagtccta 20 <210> 2886 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2886 atcttttagt ctagaacaag 20 <210> 2887 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2887 taactatctt ttagtctaga 20 <210> 2888 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2888 atctcttaac tatcttttag 20 <210> 2889 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2889 tggatatctc ttaactatct 20 <210> 2890 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2890 tttgatggat atctcttaac 20 <210> 2891 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2891 ttaattctgc ttcattaggt 20 <210> 2892 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2892 tttaattctg cttcattagg 20 <210> 2893 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2893 cagtatttaa ttctgcttca 20 <210> 2894 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2894 acagtattta attctgcttc 20 <210> 2895 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2895 tacagtattt aattctgctt 20 <210> 2896 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2896 atacagtatt taattctgct 20 <210> 2897 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2897 aatacagtat ttaattctgc 20 <210> 2898 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2898 taatacagta tttaattctg 20 <210> 2899 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2899 tttaatacag tatttaattc 20 <210> 2900 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2900 ttttaataca gtatttaatt 20 <210> 2901 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2901 ttatattttaat acagtattta 20 <210> 2902 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2902 aacttatttt aatacagtat 20 <210> 2903 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2903 gaacttattt taatacagta 20 <210> 2904 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2904 cgaacttatt ttaatacagt 20 <210> 2905 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2905 gcgaacttat tttaatacag 20 <210> 2906 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2906 agcgaactta ttttaataca 20 <210> 2907 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2907 cagcgaactt attttaatac 20 <210> 2908 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2908 acagcgaact tattttaata 20 <210> 2909 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2909 gacagcgaac ttattttaat 20 <210> 2910 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2910 agacagcgaa cttattttaa 20 <210> 2911 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2911 aaagacagcg aacttatttt 20 <210> 2912 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2912 taaagacagc gaacttattt 20 <210> 2913 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2913 gtttaaagac agcgaactta 20 <210> 2914 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2914 tgtttaaaga cagcgaactt 20 <210> 2915 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2915 ttgtttaaag acagcgaact 20 <210> 2916 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2916 tttgtttaaa gacagcgaac 20 <210> 2917 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2917 atttgtttaa agacagcgaa 20 <210> 2918 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2918 catttgttta aagacagcga 20 <210> 2919 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2919 tccatttgtt taaagacagc 20 <210> 2920 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2920 ctccatttgt ttaaagacag 20 <210> 2921 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2921 atctccattt gtttaaagac 20 <210> 2922 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2922 catctccatt tgtttaaaga 20 <210> 2923 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2923 tcatctccat ttgtttaaag 20 <210> 2924 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2924 gtcatctcca tttgtttaaa 20 <210> 2925 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2925 tagtcatctc catttgttta 20 <210> 2926 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2926 gtagtcatct ccatttgttt 20 <210> 2927 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2927 agtagtcatc tccatttgtt 20 <210> 2928 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2928 tagtagtcat ctccatttgt 20 <210> 2929 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2929 ttagtagtca tctccatttg 20 <210> 2930 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2930 cttagtagtc atctccattt 20 <210> 2931 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2931 gtgacttagt agtcatctcc 20 <210> 2932 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2932 tgtgacttag tagtcatctc 20 <210> 2933 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2933 atgtgactta gtagtcatct 20 <210> 2934 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2934 aatgtgactt agtagtcatc 20 <210> 2935 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2935 caatgtgact tagtagtcat 20 <210> 2936 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2936 tcaatgtgac ttagtagtca 20 <210> 2937 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2937 agtcaatgtg acttagtagt 20 <210> 2938 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2938 ttaaagtcaa tgtgacttag 20 <210> 2939 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2939 gttaaagtca atgtgactta 20 <210> 2940 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2940 tgttaaagtc aatgtgactt 20 <210> 2941 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2941 atgttaaagt caatgtgact 20 <210> 2942 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2942 catgttaaag tcaatgtgac 20 <210> 2943 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2943 ctcatgttaa agtcaatgtg 20 <210> 2944 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2944 cctcatgtta aagtcaatgt 20 <210> 2945 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2945 acctcatgtt aaagtcaatg 20 <210> 2946 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2946 tacctcatgt taaagtcaat 20 <210> 2947 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2947 atacctcatg ttaaagtcaa 20 <210> 2948 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2948 gatacctcat gttaaagtca 20 <210> 2949 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2949 tgatacctca tgttaaagtc 20 <210> 2950 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2950 gtgatacctc atgttaaagt 20 <210> 2951 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2951 agtgatacct catgttaaag 20 <210> 2952 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2952 tagtgatacc tcatgttaaa 20 <210> 2953 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2953 atagtgatac ctcatgttaa 20 <210> 2954 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2954 tatagtgata cctcatgtta 20 <210> 2955 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2955 gtatagtgat acctcatgtt 20 <210> 2956 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2956 ggtatagtga tacctcatgt 20 <210> 2957 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2957 ataaggtata gtgatacctc 20 <210> 2958 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2958 aataaggtat agtgatacct 20 <210> 2959 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2959 taatgtttaa attattgcct 20 <210> 2960 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2960 ggttaatgtt taaattattg 20 <210> 2961 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2961 aggttaatgt ttaaattatt 20 <210> 2962 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2962 gaggttaatg tttaaattat 20 <210> 2963 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2963 tgaggttaat gtttaaatta 20 <210> 2964 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2964 aatgaggtta atgtttaaat 20 <210> 2965 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2965 gaatgaggtt aatgtttaaa 20 <210> 2966 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2966 ggaatgaggt taatgtttaa 20 <210> 2967 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2967 tggaatgagg ttaatgttta 20 <210> 2968 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2968 ttggaatgag gttaatgttt 20 <210> 2969 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2969 cttggaatga ggttaatgtt 20 <210> 2970 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2970 taacttggaa tgaggttaat 20 <210> 2971 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2971 ttaacttgga atgaggttaa 20 <210> 2972 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2972 attaacttgg aatgaggtta 20 <210> 2973 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2973 cattaacttg gaatgaggtt 20 <210> 2974 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2974 acattaactt ggaatgaggt 20 <210> 2975 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2975 accacattaa cttggaatga 20 <210> 2976 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2976 gaccacatta acttggaatg 20 <210> 2977 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2977 agaccacatt aacttggaat 20 <210> 2978 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2978 tagaccacat taacttggaa 20 <210> 2979 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2979 ttagaccaca ttaacttgga 20 <210> 2980 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2980 attagaccac attaacttgg 20 <210> 2981 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2981 tattagacca cattaacttg 20 <210> 2982 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2982 ttattagacc acattaactt 20 <210> 2983 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2983 attattagac cacattaact 20 <210> 2984 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2984 gattattaga ccacattaac 20 <210> 2985 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2985 aataccagat tattagacca 20 <210> 2986 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2986 taataccaga ttattagacc 20 <210> 2987 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2987 tttaatacca gattattaga 20 <210> 2988 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2988 atttaatacc agattattag 20 <210> 2989 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2989 gattaatac cagattatta 20 <210> 2990 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2990 taaggattta ataccagatt 20 <210> 2991 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2991 tttctcttaa ggatttaata 20 <210> 2992 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2992 ctttctctta aggatttaat 20 <210> 2993 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2993 agctttctct taaggattta 20 <210> 2994 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2994 aagctttctc ttaaggattt 20 <210> 2995 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2995 tcaagctttc tcttaaggat 20 <210> 2996 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2996 ttctcaagct ttctcttaag 20 <210> 2997 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2997 tttctcaagc tttctcttaa 20 <210> 2998 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2998 tctatttctc aagctttctc 20 <210> 2999 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 2999 atctatttct caagctttct 20 <210> 3000 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3000 aatctatttc tcaagctttc 20 <210> 3001 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3001 aaatctattt ctcaagcttt 20 <210> 3002 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3002 aaaatctatt tctcaagctt 20 <210> 3003 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3003 gataaaaaaa atctatttct 20 <210> 3004 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3004 tagacagtga ctttaagata 20 <210> 3005 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3005 atagacagtg actttaagat 20 <210> 3006 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3006 aataagacagt gactttaaga 20 <210> 3007 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3007 aaataagacag tgactttaag 20 <210> 3008 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3008 taaatagaca gtgactttaa 20 <210> 3009 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3009 ttaaatagac agtgacttta 20 <210> 3010 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3010 cttaaataga cagtgacttt 20 <210> 3011 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3011 tcttaaatag acagtgactt 20 <210> 3012 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3012 atcttaaata gacagtgact 20 <210> 3013 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3013 aatcttaaat agacagtgac 20 <210> 3014 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3014 taatcttaaa tagacagtga 20 <210> 3015 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3015 ttaatcttaa atagacagtg 20 <210> 3016 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3016 tttaatctta aatagacagt 20 <210> 3017 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3017 gtttaatctt aaataagacag 20 <210> 3018 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3018 tgtttaatct taaatagaca 20 <210> 3019 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3019 atgtttaatc ttaaatatagac 20 <210> 3020 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3020 ttgtatgttt aatcttaaat 20 <210> 3021 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3021 attgtatgtt taatcttaaa 20 <210> 3022 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3022 gattgtatgt ttaatcttaa 20 <210> 3023 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3023 tgattgtatg tttaatctta 20 <210> 3024 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3024 tatgtgattg tatgtttaat 20 <210> 3025 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3025 gttatgtgat tgtatgttta 20 <210> 3026 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3026 taaggttatg tgattgtatg 20 <210> 3027 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3027 ttaaggttat gtgattgtat 20 <210> 3028 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3028 ttctttaagg ttatgtgatt 20 <210> 3029 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3029 tctggaagca gaccta 16 <210> 3030 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3030 cttctggaag cagacc 16 <210> 3031 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3031 aaataaggta tagtga 16 <210> 3032 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3032 tagtattaag tgttaa 16 <210> 3033 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3033 tcatagtatt aagtgt 16 <210> 3034 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3034 agattccttt acaatt 16 <210> 3035 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3035 acaagattcc tttaca 16 <210> 3036 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3036 ctgacaagat tccttt 16 <210> 3037 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3037 tgtaatctga caagat 16 <210> 3038 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3038 tactgtaatc tgacaa 16 <210> 3039 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3039 tcttactgta atctga 16 <210> 3040 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3040 cattcttact gtaatc 16 <210> 3041 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3041 gttcattctt actgta 16 <210> 3042 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3042 atatgttcat tcttac 16 <210> 3043 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3043 gccacaaata tgttca 16 <210> 3044 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3044 gatgccacaa atatgt 16 <210> 3045 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3045 ctttaactcg atgcca 16 <210> 3046 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3046 aaactttaac tcgatg 16 <210> 3047 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3047 tataaacttt aactcg 16 <210> 3048 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3048 cacagcatat ttaggg 16 <210> 3049 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3049 tagaatcaca gcatat 16 <210> 3050 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3050 tattagaatc acagca 16 <210> 3051 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3051 aatgtattag aatcac 16 <210> 3052 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3052 acgaatgtat tagaat 16 <210> 3053 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3053 tacacgaatg tattag 16 <210> 3054 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3054 acctacacga atgtat 16 <210> 3055 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3055 aaaacctaca cgaatg 16 <210> 3056 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3056 ttgaaaacct acacga 16 <210> 3057 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3057 tacttgaaaa cctaca 16 <210> 3058 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3058 gtttatttct acttga 16 <210> 3059 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3059 gaggtttatt tctact 16 <210> 3060 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3060 tacgaggttt atttct 16 <210> 3061 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3061 tgttacgagg tttatt 16 <210> 3062 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3062 acttgttacg aggttt 16 <210> 3063 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3063 cagtaacttg ttacga 16 <210> 3064 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3064 gttcagtaac ttgtta 16 <210> 3065 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3065 tcaggctgtt taaacg 16 <210> 3066 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3066 ttgtcaggct gtttaa 16 <210> 3067 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3067 tatacatgct tgtcag 16 <210> 3068 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3068 gtctttgttt attgaa 16 <210> 3069 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3069 tgggtctttg tttatt 16 <210> 3070 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3070 gactgggtct ttgttt 16 <210> 3071 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3071 ataatttag gactgg 16 <210> 3072 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3072 tctataattt agggac 16 <210> 3073 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3073 cgataaacat gcaaga 16 <210> 3074 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3074 tgtcgataaa catgca 16 <210> 3075 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3075 tgatgtcgat aaacat 16 <210> 3076 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3076 ttgtgatgtc gataaa 16 <210> 3077 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3077 ctgttgtgat gtcgat 16 <210> 3078 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3078 gatctgttgt gatgtc 16 <210> 3079 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3079 agggatctgt tgtgat 16 <210> 3080 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3080 tttagggatc tgttgt 16 <210> 3081 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3081 ggatttaggg atctgt 16 <210> 3082 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3082 gattaggga tttagg 16 <210> 3083 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3083 tctttaggga tttagg 16 <210> 3084 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3084 taatctttag ggattt 16 <210> 3085 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3085 atctaatctt taggga 16 <210> 3086 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3086 tgtatctaat ctttag 16 <210> 3087 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3087 aaatttgtat ctaatc 16 <210> 3088 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3088 gtaaaaaatt tgtatc 16 <210> 3089 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3089 gtggtaaaaa atttgt 16 <210> 3090 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3090 gatactgtgg taaaaa 16 <210> 3091 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3091 agtgatactg tggtaa 16 <210> 3092 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3092 acaagtgata ctgtgg 16 <210> 3093 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3093 ctgacaagtg atactg 16 <210> 3094 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3094 attctgacaa gtgata 16 <210> 3095 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3095 taaattctga caagtg 16 <210> 3096 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3096 tactggcagt tttaaa 16 <210> 3097 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3097 tcttactggc agtttt 16 <210> 3098 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3098 atttcttact ggcagt 16 <210> 3099 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3099 aaaatttctt actggc 16 <210> 3100 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3100 aacaaatggg tttaat 16 <210> 3101 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3101 gaatatttgc aagtct 16 <210> 3102 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3102 tcattggtag aggaat 16 <210> 3103 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3103 atattttaaa gtctcg 16 <210> 3104 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3104 gttacattat tataga 16 <210> 3105 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3105 gtgaaatgtg ttacat 16 <210> 3106 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3106 tcaccagtga aatgtg 16 <210> 3107 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3107 catgtttcac cagtga 16 <210> 3108 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3108 acaagacatg tttcac 16 <210> 3109 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3109 catatgacaa gacatg 16 <210> 3110 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3110 ctataatgca tatgac 16 <210> 3111 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3111 tcctttctat aatgca 16 <210> 3112 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3112 tgattatcct ttctat 16 <210> 3113 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3113 taactgaaag tctgat 16 <210> 3114 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3114 gtgcacaaaa atgtta 16 <210> 3115 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3115 agctatgtgc acaaaa 16 <210> 3116 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3116 gaagatagct atgtgc 16 <210> 3117 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3117 tttattgaag atagct 16 <210> 3118 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3118 tcattttagt gtatct 16 <210> 3119 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3119 ccttgatcat tttagt 16 <210> 3120 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3120 tgaatccctt gatcat 16 <210> 3121 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3121 gttgtttagt cttgaa 16 <210> 3122 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3122 aattgagttg tttagt 16 <210> 3123 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3123 gcaactaatt gagttg 16 <210> 3124 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3124 attggtgcaa ctaatt 16 <210> 3125 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3125 gttttttatt ggtgca 16 <210> 3126 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3126 ggacactgac agtttt 16 <210> 3127 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3127 caggttggac actgac 16 <210> 3128 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3128 taagtacagg ttggac 16 <210> 3129 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3129 agttattaag tacagg 16 <210> 3130 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3130 tctgtgagtt attaag 16 <210> 3131 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3131 accaaaattc tcctga 16 <210> 3132 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3132 acctgaataa ccctct 16 <210> 3133 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3133 ggtatcagaa aaagat 16 <210> 3134 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3134 agtattggta tcagaa 16 <210> 3135 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3135 ggaagatact ttgaag 16 <210> 3136 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3136 aatgtgggaa gatact 16 <210> 3137 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3137 cagataatag ctaata 16 <210> 3138 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3138 tcattgcaga taatag 16 <210> 3139 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3139 gattcggatt tttaaa 16 <210> 3140 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3140 atttgggatt cggatt 16 <210> 3141 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3141 acgcttattt gggatt 16 <210> 3142 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3142 tctagagaga aaacgc 16 <210> 3143 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3143 agttaagagg ttttcg 16 <210> 3144 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3144 cattatagtt aagagg 16 <210> 3145 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3145 cactttcatt attagtt 16 <210> 3146 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3146 tagaatgaac actttc 16 <210> 3147 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3147 ttgaactaga atgaac 16 <210> 3148 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3148 acctgattga actaga 16 <210> 3149 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3149 taaaatacct gattga 16 <210> 3150 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3150 tagaggtaaa atacct 16 <210> 3151 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3151 gaagattaga ggtaaa 16 <210> 3152 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3152 tctgaggaag attaga 16 <210> 3153 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3153 tatacactac caaaaa 16 <210> 3154 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3154 ataatctata cactac 16 <210> 3155 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3155 taagtcccaa ttttaa 16 <210> 3156 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3156 ccagttttaa ataatc 16 <210> 3157 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3157 tgtatcccag ttttaa 16 <210> 3158 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3158 gttttagatg catgta 16 <210> 3159 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3159 tacagtgttt tagatg 16 <210> 3160 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3160 ttccccgtaa gtttat 16 <210> 3161 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3161 ctgtatttcc ccgtaa 16 <210> 3162 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3162 tagttactgt tactgt 16 <210> 3163 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3163 cgtatgtagt tactgt 16 <210> 3164 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3164 aatgggtaca gactcg 16 <210> 3165 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3165 gcaatttaat gggtac 16 <210> 3166 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3166 gatagatatg caattt 16 <210> 3167 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3167 aaaggagata gatatg 16 <210> 3168 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3168 cctcctaaag gagata 16 <210> 3169 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3169 caccagcctc ctaaag 16 <210> 3170 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3170 tttgtgatcc caagta 16 <210> 3171 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3171 gctttgtgat cccaag 16 <210> 3172 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3172 ttgctttgtg atccca 16 <210> 3173 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3173 ttttgctttg tgatcc 16 <210> 3174 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3174 ccttttgctt tgtgat 16 <210> 3175 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3175 gtccttttgc tttgtg 16 <210> 3176 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3176 gtgtcctttt gctttg 16 <210> 3177 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3177 gaaatgtaaa cggtat 16 <210> 3178 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3178 ttgagaaatg taaacg 16 <210> 3179 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3179 tgattgagaa atgtaa 16 <210> 3180 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3180 tttgattgag aaatgt 16 <210> 3181 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3181 aagaattttg attgag 16 <210> 3182 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3182 ataagaattt tgattg 16 <210> 3183 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3183 caaatagtat tataag 16 <210> 3184 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3184 cccacatcac aaaatt 16 <210> 3185 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3185 gattcccaca tcacaa 16 <210> 3186 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3186 ttgattccca catcac 16 <210> 3187 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3187 aattgattcc cacatc 16 <210> 3188 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3188 aaaattgatt cccaca 16 <210> 3189 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3189 ctaaaattga ttccca 16 <210> 3190 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3190 catctaaaat tgattc 16 <210> 3191 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3191 accatctaaa attgat 16 <210> 3192 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3192 tgaccatcta aaattg 16 <210> 3193 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3193 tgtgaccatc taaaat 16 <210> 3194 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3194 attgtgacca tctaaa 16 <210> 3195 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3195 agattgtgac catcta 16 <210> 3196 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3196 ctagattgtg accatc 16 <210> 3197 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3197 atctagattg tgacca 16 <210> 3198 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3198 taatctagat tgtgac 16 <210> 3199 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3199 tataatctag attgtg 16 <210> 3200 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3200 attataatct agattg 16 <210> 3201 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3201 tgattataat ctagat 16 <210> 3202 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3202 attgattata atctag 16 <210> 3203 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3203 ctattgatta taatct 16 <210> 3204 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3204 acctattgat tataat 16 <210> 3205 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3205 tcacctattg attata 16 <210> 3206 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3206 gttcacctat tgatta 16 <210> 3207 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3207 aagttcacct attgat 16 <210> 3208 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3208 ataagttcac ctattg 16 <210> 3209 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3209 taataagttc acctat 16 <210> 3210 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3210 tttaataagt tcacct 16 <210> 3211 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3211 tattataataa gttcac 16 <210> 3212 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3212 gttatttaat aagttc 16 <210> 3213 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3213 catatgatgc ctttta 16 <210> 3214 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3214 tattagctca tatgat 16 <210> 3215 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3215 gatattagct catatg 16 <210> 3216 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3216 gtgatattag ctcata 16 <210> 3217 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3217 agttgtgata ttagct 16 <210> 3218 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3218 aaagttgtga tattag 16 <210> 3219 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3219 ggaaagttgt gatatt 16 <210> 3220 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3220 tgggaaagtt gtgata 16 <210> 3221 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3221 aaactgggaa agttgt 16 <210> 3222 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3222 ttaaactggg aaagtt 16 <210> 3223 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3223 gttttttaaa ctggga 16 <210> 3224 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3224 tagtttttta aactgg 16 <210> 3225 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3225 gagtactagt ttttta 16 <210> 3226 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3226 aagagtacta gttttt 16 <210> 3227 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3227 acaagagtac tagttt 16 <210> 3228 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3228 taacaagagt actagt 16 <210> 3229 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3229 tttaacaaga gtacta 16 <210> 3230 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3230 gttttaacaa gagtac 16 <210> 3231 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3231 gagttttaac aagagt 16 <210> 3232 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3232 tagagtttta acaaga 16 <210> 3233 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3233 gtttagagtt ttaaca 16 <210> 3234 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3234 caagtttaga gtttta 16 <210> 3235 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3235 gtcaagttta gagttt 16 <210> 3236 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3236 tagtcaagtt tagagt 16 <210> 3237 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3237 tttagtcaag tttaga 16 <210> 3238 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3238 tatttagtca agttta 16 <210> 3239 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3239 tgtatttagt caagtt 16 <210> 3240 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3240 tctgtatttta gtcaag 16 <210> 3241 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3241 cctctgtatt tagtca 16 <210> 3242 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3242 gtcctctgta tttagt 16 <210> 3243 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3243 cagtcctctg tattta 16 <210> 3244 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3244 accagtcctc tgtatt 16 <210> 3245 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3245 ttaccagtcc tctgta 16 <210> 3246 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3246 aattaccagt cctctg 16 <210> 3247 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3247 acaattacca gtcctc 16 <210> 3248 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3248 tctgacaaga ttcctt 16 <210> 3249 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3249 atctgacaag attcct 16 <210> 3250 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3250 taatctgaca agattc 16 <210> 3251 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3251 gtaatctgac aagatt 16 <210> 3252 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3252 cttgtcaggc tgttta 16 <210> 3253 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3253 gcttgtcagg ctgttt 16 <210> 3254 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3254 atgcttgtca ggctgt 16 <210> 3255 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3255 tacatgcttg tcaggc 16 <210> 3256 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3256 atacatgctt gtcagg 16 <210> 3257 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3257 aaaggtaggt caccat 16 <210> 3258 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3258 caaaggtagg tcacca 16 <210> 3259 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3259 gacaaaggta ggtcac 16 <210> 3260 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3260 tgacaaaggt aggtca 16 <210> 3261 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3261 tctgacatag cttttt 16 <210> 3262 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3262 attctgacat agcttt 16 <210> 3263 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3263 gattctgaca tagctt 16 <210> 3264 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3264 ggattctgac atagct 16 <210> 3265 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3265 atggattctg acatag 16 <210> 3266 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3266 catggattct gacata 16 <210> 3267 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3267 acatggattc tgacat 16 <210> 3268 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3268 tacatggatt ctgaca 16 <210> 3269 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3269 atacatggat tctgac 16 <210> 3270 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3270 tttagcagca ctacta 16 <210> 3271 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3271 ttttagcagc actact 16 <210> 3272 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3272 cttttagcag cactac 16 <210> 3273 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3273 aaaacctttt agcagc 16 <210> 3274 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3274 gataaaaaac ctttta 16 <210> 3275 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3275 tgataaaaaa cctttt 16 <210> 3276 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3276 agatgttggc aggttg 16 <210> 3277 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3277 tagatgttgg caggtt 16 <210> 3278 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3278 gtagatgttg gcaggt 16 <210> 3279 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3279 tgtagatgtt ggcagg 16 <210> 3280 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3280 ctgtagatgt tggcag 16 <210> 3281 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3281 tatctgtaga tgttgg 16 <210> 3282 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3282 atatctgtag atgttg 16 <210> 3283 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3283 cattctgta gatgtt 16 <210> 3284 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3284 tttgaaccag gctttc 16 <210> 3285 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3285 aatttgaacc aggctt 16 <210> 3286 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3286 cataatttga accagg 16 <210> 3287 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3287 acataatttg aaccag 16 <210> 3288 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3288 tacataattt gaacca 16 <210> 3289 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3289 atacataatt tgaacc 16 <210> 3290 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3290 acattggtcg gaaaac 16 <210> 3291 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3291 gacattggtc ggaaaa 16 <210> 3292 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3292 agacattggt cggaaa 16 <210> 3293 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3293 cagacattgg tcggaa 16 <210> 3294 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3294 gcagacattg gtcgga 16 <210> 3295 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3295 aagcagacat tggtcg 16 <210> 3296 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3296 tgtacagatt acctgt 16 <210> 3297 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3297 ttgtacagat tacctg 16 <210> 3298 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3298 attgtacaga ttacct 16 <210> 3299 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3299 gattgtacag attacc 16 <210> 3300 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3300 agattgtaca gattac 16 <210> 3301 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3301 tcagattgta cagatt 16 <210> 3302 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3302 ttcagattgt acagat 16 <210> 3303 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3303 attcagattg tacaga 16 <210> 3304 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3304 tattcagatt gtacag 16 <210> 3305 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3305 ttattcagat tgtaca 16 <210> 3306 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3306 taggtatgtc ttttat 16 <210> 3307 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3307 tgtcttaggt atgtct 16 <210> 3308 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3308 attgtcttag gtatgt 16 <210> 3309 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3309 gattgtctta ggtatg 16 <210> 3310 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3310 ttcttagatg gcgtgt 16 <210> 3311 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3311 atttttctta gatggc 16 <210> 3312 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3312 catttttctt agatgg 16 <210> 3313 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3313 gcatttttct tagatg 16 <210> 3314 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3314 ataagtccca atttta 16 <210> 3315 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3315 tataagtccc aatttt 16 <210> 3316 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3316 gtataagtcc caattt 16 <210> 3317 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3317 tgtataagtc ccaatt 16 <210> 3318 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3318 ctgtataagt cccaat 16 <210> 3319 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3319 taatctgtat aagtcc 16 <210> 3320 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3320 ataatctgta taagtc 16 <210> 3321 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3321 aataatctgt ataagt 16 <210> 3322 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3322 tgcatgtatc ccagtt 16 <210> 3323 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3323 tagatgcatg tatccc 16 <210> 3324 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3324 ttagatgcat gtatcc 16 <210> 3325 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3325 tttagatgca tgtatc 16 <210> 3326 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3326 gtggaactgt tttctt 16 <210> 3327 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3327 acgtggaact gttttc 16 <210> 3328 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3328 ttgatcaatt ctggag 16 <210> 3329 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3329 tcttgatcaa ttctgg 16 <210> 3330 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3330 ggctctggag atagag 16 <210> 3331 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3331 ttggctctgg agatag 16 <210> 3332 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3332 gattttggct ctggag 16 <210> 3333 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3333 tcttgatttt ggctct 16 <210> 3334 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3334 aatcttgatt ttggct 16 <210> 3335 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3335 caaatcttga ttttgg 16 <210> 3336 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3336 gcagcgatag atcata 16 <210> 3337 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3337 ttgcagcgat agatca 16 <210> 3338 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3338 tggtttgcag cgatag 16 <210> 3339 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3339 tttgatttca ctggtt 16 <210> 3340 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3340 tctttgattt cactgg 16 <210> 3341 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3341 agttcttctc agttcc 16 <210> 3342 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3342 ttagttagtt gctctt 16 <210> 3343 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3343 agttagttag ttgctc 16 <210> 3344 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3344 gtgctcttgg cttgga 16 <210> 3345 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3345 tggtgctctt ggcttg 16 <210> 3346 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3346 tatgttcacc tctgtt 16 <210> 3347 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3347 tgtatgttca cctctg 16 <210> 3348 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3348 acactcatca tgccac 16 <210> 3349 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3349 ccacactcat catgcc 16 <210> 3350 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3350 ttagattttg ctcttg 16 <210> 3351 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3351 atagagtata accttc 16 <210> 3352 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3352 aatctgttgg atggat 16 <210> 3353 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3353 tgaatctgtt ggatgg 16 <210> 3354 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3354 ttcattcaaa gctttc 16 <210> 3355 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3355 agttcattca aagctt 16 <210> 3356 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3356 tcagttcatt caaagc 16 <210> 3357 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3357 gattattaga ccacat 16 <210> 3358 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3358 aataagacagt gacttt 16 <210> 3359 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3359 agaaatgtaa acggta 16 <210> 3360 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3360 tcatatgatg cctttt 16 <210> 3361 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3361 attagctcat atgatg 16 <210> 3362 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3362 tgtgatatta gctcat 16 <210> 3363 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3363 gttgtgatat tagctc 16 <210> 3364 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3364 tactctgtgc tgacga 16 <210> 3365 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3365 catactctgt gctgac 16 <210> 3366 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3366 gtttaaagac agcgaa 16 <210> 3367 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3367 ttgtttaaag acagcg 16 <210> 3368 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3368 gtagtcatct ccattt 16 <210> 3369 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3369 tagtagtcat ctccat 16 <210> 3370 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3370 tcttaactct atatat 16 <210> 3371 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3371 cttcttaact ctatat 16 <210> 3372 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3372 gacttcttaa ctctat 16 <210> 3373 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3373 tagacttctt aactct 16 <210> 3374 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3374 cctagacttc ttaact 16 <210> 3375 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3375 gacctagact tcttaa 16 <210> 3376 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3376 cagacctaga cttctt 16 <210> 3377 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3377 agcagaccta gacttc 16 <210> 3378 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3378 gaagcagacc tagact 16 <210> 3379 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3379 tggaagcaga cctaga 16 <210> 3380 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3380 ctttaacaaa tgggtt 16 <210> 3381 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3381 atcctttaac aaatgg 16 <210> 3382 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3382 ctatatcctt taacaa 16 <210> 3383 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3383 gcactatatc ctttaa 16 <210> 3384 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3384 tgggcactat atcctt 16 <210> 3385 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3385 acttgggcac tatatc 16 <210> 3386 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3386 ataacttggg cactat 16 <210> 3387 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3387 catataactt gggcac 16 <210> 3388 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3388 caccatataa cttggg 16 <210> 3389 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3389 ggtcaccata taactt 16 <210> 3390 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3390 gtaggtcacc atataa 16 <210> 3391 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3391 aaggtaggtc accata 16 <210> 3392 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3392 ttgacaaagg taggtc 16 <210> 3393 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3393 gtattgacaa aggtag 16 <210> 3394 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3394 taagtattga caaagg 16 <210> 3395 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3395 tgctaagtat tgacaa 16 <210> 3396 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3396 taatgctaag tattga 16 <210> 3397 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3397 tacataatgc taagta 16 <210> 3398 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3398 ggataatttg aaatac 16 <210> 3399 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3399 tattggataa tttgaa 16 <210> 3400 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3400 gtattattgga taattt 16 <210> 3401 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3401 catgtatatt ggataa 16 <210> 3402 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3402 cttttatata tgtgac 16 <210> 3403 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3403 gatcatacat atcttt 16 <210> 3404 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3404 atagatcata catatc 16 <210> 3405 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3405 cacatagatc atacat 16 <210> 3406 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3406 attcacatag atcata 16 <210> 3407 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3407 aggattcaca tagatc 16 <210> 3408 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3408 cttaggattc acatag 16 <210> 3409 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3409 ttacttagga ttcaca 16 <210> 3410 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3410 tatttactta ggattc 16 <210> 3411 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3411 cctgaaaatt atagat 16 <210> 3412 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3412 ggtcctgaaa attata 16 <210> 3413 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3413 tgtggtcctg aaaatt 16 <210> 3414 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3414 gtctgtggtc ctgaaa 16 <210> 3415 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3415 ttagtctgtg gtcctg 16 <210> 3416 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3416 agcttagtct gtggtc 16 <210> 3417 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3417 gacagcttag tctgtg 16 <210> 3418 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3418 ttcgacagct tagtct 16 <210> 3419 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3419 aatttcgaca gcttag 16 <210> 3420 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3420 gttaatttcg acagct 16 <210> 3421 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3421 cctaaaaaaa tcagcg 16 <210> 3422 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3422 ggccctaaaa aaatca 16 <210> 3423 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3423 ttctggccct aaaaaa 16 <210> 3424 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3424 gtattctggc cctaaa 16 <210> 3425 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3425 ttggtattct ggccct 16 <210> 3426 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3426 attttggtat tctggc 16 <210> 3427 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3427 gccattttgg tattct 16 <210> 3428 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3428 ggagccattt tggtat 16 <210> 3429 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3429 agaggagcca ttttgg 16 <210> 3430 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3430 aagagaggag ccattt 16 <210> 3431 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3431 attgtccaat tttggg 16 <210> 3432 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3432 gaaattgtcc aatttt 16 <210> 3433 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3433 tttgaaattg tccaat 16 <210> 3434 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3434 gcatttgaaa ttgtcc 16 <210> 3435 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3435 gcaactcata tattata 16 <210> 3436 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3436 gaagcaactc atatat 16 <210> 3437 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3437 gaggaagcaa ctcata 16 <210> 3438 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3438 atagaggaag caactc 16 <210> 3439 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3439 caaatagagg aagcaa 16 <210> 3440 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3440 aaccaaatag aggaag 16 <210> 3441 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3441 ggaaaccaaa tagagg 16 <210> 3442 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3442 ctttaagtga agttac 16 <210> 3443 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3443 tacttacttt aagtga 16 <210> 3444 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3444 gaaccctctt tatttt 16 <210> 3445 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3445 aaacatgaac cctctt 16 <210> 3446 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3446 gatccacatt gaaaac 16 <210> 3447 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3447 catgccttag aaatat 16 <210> 3448 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3448 aaatggcatg ccttag 16 <210> 3449 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3449 gtatttcaaa tggcat 16 <210> 3450 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3450 gcaacaaagt atttca 16 <210> 3451 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3451 gtatttcaac aatgca 16 <210> 3452 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3452 ataacattag ggaaac 16 <210> 3453 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3453 tcatatataa cattag 16 <210> 3454 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3454 gtggttttga gcaaag 16 <210> 3455 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3455 ctattgtgtg gttttg 16 <210> 3456 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3456 ggaaagctat tgtgtg 16 <210> 3457 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3457 tatgagtgaa atggaa 16 <210> 3458 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3458 agccaatatg agtgaa 16 <210> 3459 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3459 ctaaagagcc aatatg 16 <210> 3460 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3460 ggtaatcttg gtctaa 16 <210> 3461 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3461 gatgacgaag ggttgg 16 <210> 3462 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3462 cagtgagatg acgaag 16 <210> 3463 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3463 tgaagtcagt gagatg 16 <210> 3464 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3464 aggaggtgaa gtcagt 16 <210> 3465 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3465 gagtagagga ggtgaa 16 <210> 3466 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3466 taactagagt agagga 16 <210> 3467 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3467 tcagaataac tagagt 16 <210> 3468 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3468 aagcggtcag aataac 16 <210> 3469 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3469 ctggtaaagc ggtcag 16 <210> 3470 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3470 tgaatactgg taaagc 16 <210> 3471 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3471 tgtgtttgaa tactgg 16 <210> 3472 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3472 gtttgatgtg tttgaa 16 <210> 3473 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3473 cagtatgttt gatgtg 16 <210> 3474 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3474 aggtggcagt atgttt 16 <210> 3475 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3475 gctttgaggt ggcagt 16 <210> 3476 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3476 gggcaaaggc tttgag 16 <210> 3477 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3477 caacaagggc aaaggc 16 <210> 3478 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3478 gaggaaacaa caaggg 16 <210> 3479 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3479 ccagttagag gaaaca 16 <210> 3480 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3480 ccagggcaga agagcg 16 <210> 3481 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3481 tagataccag ggcaga 16 <210> 3482 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3482 cagagagtgg gccacg 16 <210> 3483 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3483 ggaaatcaga gagtgg 16 <210> 3484 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3484 cctaagggaa atcaga 16 <210> 3485 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3485 aacgacccta agggaa 16 <210> 3486 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3486 tttgataacg acccta 16 <210> 3487 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3487 tttttgtttg ataacg 16 <210> 3488 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3488 cattgggaat tttttg 16 <210> 3489 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3489 agtcttcatt gggaat 16 <210> 3490 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3490 cttgtaagtc ttcatt 16 <210> 3491 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3491 agtgaccttg taagtc 16 <210> 3492 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3492 tggttaagtg accttg 16 <210> 3493 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3493 gatttttggt taagtg 16 <210> 3494 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3494 ggttgtgatt tttggt 16 <210> 3495 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3495 ccaggcggtt gtgatt 16 <210> 3496 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3496 atgggaccag gcggtt 16 <210> 3497 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3497 aagttttcag ggatgg 16 <210> 3498 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3498 aagtagaagt tttcag 16 <210> 3499 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3499 ctaaggaagt agaagt 16 <210> 3500 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3500 aagtagctaa ggaagt 16 <210> 3501 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3501 ggagaaaagt agctaa 16 <210> 3502 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3502 tgtgcaggag aaaagt 16 <210> 3503 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3503 ggtgagtgtg caggag 16 <210> 3504 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3504 aataaaggtg agtgtg 16 <210> 3505 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3505 tgcaggaata gaagag 16 <210> 3506 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3506 ttttagtgca ggaata 16 <210> 3507 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3507 tattcacaga gcttac 16 <210> 3508 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3508 tccctgtatt cacaga 16 <210> 3509 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3509 gaaaaaatcc ctgtat 16 <210> 3510 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3510 tatgaagata atggaa 16 <210> 3511 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3511 ggagtata caaata 16 <210> 3512 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3512 tattctggag tatata 16 <210> 3513 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3513 attctatatt ctggag 16 <210> 3514 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3514 catacagtat tctata 16 <210> 3515 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3515 gtgtgccata cagtat 16 <210> 3516 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3516 agaaatgcct actgtg 16 <210> 3517 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3517 attcaacaga aatgcc 16 <210> 3518 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3518 gaatatgaca ttacat 16 <210> 3519 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3519 ctgtgtgaat atgaca 16 <210> 3520 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3520 acgcttctgt gtgaat 16 <210> 3521 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3521 tagcacacgc ttctgt 16 <210> 3522 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3522 taatcatagc acacgc 16 <210> 3523 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3523 ccaagtaata ataatc 16 <210> 3524 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3524 agtaatccaa gtaata 16 <210> 3525 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3525 atttctagta atccaa 16 <210> 3526 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3526 cacactattt ctagta 16 <210> 3527 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3527 atgaggcaca ctattt 16 <210> 3528 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3528 ttaattatga ggcaca 16 <210> 3529 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3529 tgacctttaa ttatga 16 <210> 3530 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3530 gcaatttatt gaatga 16 <210> 3531 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3531 gggtttgcaa tttatt 16 <210> 3532 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3532 tgtgttgggt ttgcaa 16 <210> 3533 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3533 tttaagtgtg ttgggt 16 <210> 3534 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3534 gtttagcagt aacatt 16 <210> 3535 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3535 attcagtagt ttatcg 16 <210> 3536 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3536 ctatatattc agtagt 16 <210> 3537 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3537 gcttactttc tatata 16 <210> 3538 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3538 agtttgtttg cttact 16 <210> 3539 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3539 ttggcaagtt tgtttg 16 <210> 3540 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3540 ggcaggttgg caagtt 16 <210> 3541 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3541 gatgttggca ggttgg 16 <210> 3542 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3542 aacatatctg tagatg 16 <210> 3543 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3543 cctgtaaaca tatctg 16 <210> 3544 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3544 ttttgacctg taaaca 16 <210> 3545 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3545 gataattttt gacctg 16 <210> 3546 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3546 tcttgataat ttgata 16 <210> 3547 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3547 aggctttctt gataat 16 <210> 3548 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3548 tgaaccaggc tttctt 16 <210> 3549 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3549 gataaagaca taatac 16 <210> 3550 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3550 acctgtgata aagaca 16 <210> 3551 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3551 cttcagacct gtgata 16 <210> 3552 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3552 actgatcttc agacct 16 <210> 3553 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3553 ggtcttactg atcttc 16 <210> 3554 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3554 gttttaggtc ttactg 16 <210> 3555 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3555 gttcagattt taagtt 16 <210> 3556 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3556 atattctgtt cagatt 16 <210> 3557 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3557 atattgtaat gtattc 16 <210> 3558 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3558 cttagaatat tgtaat 16 <210> 3559 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3559 gctttgctta gaatat 16 <210> 3560 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3560 gagactgctt tgctta 16 <210> 3561 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3561 aaagtagaga ctgctt 16 <210> 3562 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3562 aggccaaaag tagaga 16 <210> 3563 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3563 tcggaaaaca gagcaa 16 <210> 3564 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3564 cattggtcgg aaaaca 16 <210> 3565 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3565 agcaaggcaa aaaagc 16 <210> 3566 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3566 gacattattt aataag 16 <210> 3567 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3567 atcagggaca ttattt 16 <210> 3568 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3568 tattaatca gggaca 16 <210> 3569 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3569 attacctgtt ctcaaa 16 <210> 3570 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3570 gtacagatta cctgtt 16 <210> 3571 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3571 gttattcaga ttgtac 16 <210> 3572 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3572 aacagtgtta ttcaga 16 <210> 3573 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3573 tagataaaca gtgtta 16 <210> 3574 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3574 tgatatttag ataaac 16 <210> 3575 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3575 ggtgtttgat atttag 16 <210> 3576 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3576 tataacggtg tttgat 16 <210> 3577 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3577 taatgttata acggtg 16 <210> 3578 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3578 agttcataat gttata 16 <210> 3579 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3579 gtctttcagt tcataa 16 <210> 3580 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3580 acagtttgtc tttcag 16 <210> 3581 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3581 agaagtacag tttgtc 16 <210> 3582 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3582 gatgtcagaa gtacag 16 <210> 3583 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3583 agtaaggatg tcagaa 16 <210> 3584 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3584 aatctgagta aggatg 16 <210> 3585 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3585 gaatatacaa ttaggg 16 <210> 3586 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3586 tgatactgaa tataca 16 <210> 3587 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3587 ctgagctgat aaaaga 16 <210> 3588 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3588 accatcatgt tttaca 16 <210> 3589 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3589 tgtcttacca tcatgt 16 <210> 3590 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3590 ccaaagtgtc ttacca 16 <210> 3591 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3591 aacccaccaa agtgtc 16 <210> 3592 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3592 gaaggaaacc caccaa 16 <210> 3593 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3593 cttcaagaag gaaacc 16 <210> 3594 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3594 taatagcttc aagaag 16 <210> 3595 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3595 gggaatttga taataa 16 <210> 3596 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3596 agaataggga atttga 16 <210> 3597 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3597 gtcctaagaa taggga 16 <210> 3598 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3598 gaacaagtcc taagaa 16 <210> 3599 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3599 agtctagaac aagtcc 16 <210> 3600 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3600 tcttttagtc tagaac 16 <210> 3601 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3601 taactatctt ttagtc 16 <210> 3602 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3602 atctcttaac tatctt 16 <210> 3603 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3603 aactgttttc ttctgg 16 <210> 3604 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3604 tcaatttcaa gcaacg 16 <210> 3605 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3605 cttaattgtg aacatt 16 <210> 3606 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3606 agcttaattg tgaaca 16 <210> 3607 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3607 ggagcttaat tgtgaa 16 <210> 3608 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3608 aaggagctta attgtg 16 <210> 3609 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3609 agaaggagct taattg 16 <210> 3610 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3610 aaagaaggag cttaat 16 <210> 3611 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3611 ctagaggaac aataaa 16 <210> 3612 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3612 taactagagg aacaat 16 <210> 3613 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3613 aataactaga ggaaca 16 <210> 3614 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3614 ggaaataact agagga 16 <210> 3615 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3615 gaggaaataa ctagag 16 <210> 3616 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3616 tggaggaaat aactag 16 <210> 3617 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3617 tctggaggaa ataact 16 <210> 3618 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3618 caattctgga ggaaat 16 <210> 3619 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3619 atcaattctg gaggaa 16 <210> 3620 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3620 tgatcaattc tggagg 16 <210> 3621 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3621 ttgtcttgat caattc 16 <210> 3622 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3622 aattgtcttg atcaat 16 <210> 3623 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3623 tgaattgtct tgatca 16 <210> 3624 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3624 gatgaattgt cttgat 16 <210> 3625 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3625 atgatgaatt gtcttg 16 <210> 3626 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3626 caaatgatga attgtc 16 <210> 3627 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3627 atcaaatgat gaattg 16 <210> 3628 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3628 gatagagaat caaatg 16 <210> 3629 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3629 tggagataga gaatca 16 <210> 3630 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3630 tctggagata gagaat 16 <210> 3631 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3631 tttggctctg gagata 16 <210> 3632 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3632 attttggctc tggaga 16 <210> 3633 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3633 atcttgattt tggctc 16 <210> 3634 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3634 gcaaatcttg attttg 16 <210> 3635 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3635 tagcaaatct tgattt 16 <210> 3636 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3636 catagcaaat cttgat 16 <210> 3637 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3637 aacatagcaa atcttg 16 <210> 3638 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3638 ctaacatagc aaatct 16 <210> 3639 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3639 gtctaacata gcaaat 16 <210> 3640 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3640 taaaattttt acatcg 16 <210> 3641 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3641 ggctaaaatt tttaca 16 <210> 3642 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3642 ccattggcta aaattt 16 <210> 3643 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3643 ggccattggc taaaat 16 <210> 3644 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3644 gaggccattg gctaaa 16 <210> 3645 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3645 aggaggccat tggcta 16 <210> 3646 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3646 gaaggaggcc attggc 16 <210> 3647 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3647 ctgaaggagg ccattg 16 <210> 3648 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3648 aactgaagga ggccat 16 <210> 3649 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3649 ccaactgaag gaggcc 16 <210> 3650 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3650 tcccaactga aggagg 16 <210> 3651 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3651 tgtcccaact gaagga 16 <210> 3652 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3652 catgtcccaa ctgaag 16 <210> 3653 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3653 accatgtccc aactga 16 <210> 3654 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3654 agaccatgtc ccaact 16 <210> 3655 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3655 taagaccatg tcccaa 16 <210> 3656 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3656 tttaagacca tgtccc 16 <210> 3657 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3657 tctttaagac catgtc 16 <210> 3658 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3658 agtctttaag accatg 16 <210> 3659 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3659 aaagtcttta agacca 16 <210> 3660 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3660 acaaagtctt taagac 16 <210> 3661 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3661 ggacaaagtc tttaag 16 <210> 3662 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3662 atggacaaag tcttta 16 <210> 3663 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3663 ttatggacaa agtctt 16 <210> 3664 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3664 tcttatggac aaagtc 16 <210> 3665 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3665 cgtcttatgg acaaag 16 <210> 3666 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3666 ttaatttggc ccttcg 16 <210> 3667 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3667 cattaatttg gccctt 16 <210> 3668 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3668 gtcattaatt tggccc 16 <210> 3669 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3669 atgtcattaa tttggc 16 <210> 3670 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3670 tatgttgagt ttttga 16 <210> 3671 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3671 tcaaatatgt tgagtt 16 <210> 3672 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3672 gatcaaatat gttgag 16 <210> 3673 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3673 gtacaattac cagtcc 16 <210> 3674 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3674 ctgtacaatt accagt 16 <210> 3675 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3675 aactgtacaa ttacca 16 <210> 3676 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3676 agaactgtac aattac 16 <210> 3677 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3677 taagaactgt acaatt 16 <210> 3678 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3678 catttaagaa ctgtac 16 <210> 3679 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3679 tactacaaca tttaag 16 <210> 3680 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3680 ttaatactac aacatt 16 <210> 3681 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3681 ttgaaattaa tactac 16 <210> 3682 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3682 gttttgaaat taatac 16 <210> 3683 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3683 cgatttttag ttttga 16 <210> 3684 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3684 gacgattttt agtttt 16 <210> 3685 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3685 ctgacgattt ttagtt 16 <210> 3686 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3686 tgctgacgat ttttag 16 <210> 3687 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3687 tgtgctgacg attttt 16 <210> 3688 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3688 tctgtgctga cgattt 16 <210> 3689 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3689 tacacatact ctgtgc 16 <210> 3690 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3690 tttacacata ctctgt 16 <210> 3691 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3691 gatttttaca catact 16 <210> 3692 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3692 gaagcatcag tttaaa 16 <210> 3693 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3693 atgaagcatc agttta 16 <210> 3694 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3694 ttgtagcaaa atgaag 16 <210> 3695 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3695 catttactcc aaatta 16 <210> 3696 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3696 cattaggttt cataaa 16 <210> 3697 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3697 ttcattaggt ttcata 16 <210> 3698 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3698 gcttcattag gtttca 16 <210> 3699 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3699 ctgcttcatt aggttt 16 <210> 3700 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3700 ttctgcttca ttaggt 16 <210> 3701 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3701 aattctgctt cattag 16 <210> 3702 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3702 cagtatttaa ttctgc 16 <210> 3703 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3703 gaacttattt taatac 16 <210> 3704 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3704 gcgaacttat tttaat 16 <210> 3705 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3705 cagcgaactt atttta 16 <210> 3706 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3706 gacagcgaac ttattt 16 <210> 3707 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3707 aagacagcga acttat 16 <210> 3708 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3708 tttaaagaca gcgaac 16 <210> 3709 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3709 gtcatctcca tttgtt 16 <210> 3710 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3710 tagtcatctc catttg 16 <210> 3711 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3711 acttagtagt catctc 16 <210> 3712 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3712 tgacttagta gtcatc 16 <210> 3713 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3713 tgtgacttag tagtca 16 <210> 3714 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3714 aatgtgactt agtagt 16 <210> 3715 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3715 tcaatgtgac ttagta 16 <210> 3716 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3716 agtcaatgtg acttag 16 <210> 3717 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3717 aaagtcaatg tgactt 16 <210> 3718 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3718 ttaaagtcaa tgtgac 16 <210> 3719 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3719 tgttaaagtc aatgtg 16 <210> 3720 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3720 catgttaaag tcaatg 16 <210> 3721 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3721 ctcatgttaa agtcaa 16 <210> 3722 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3722 acctcatgtt aaagtc 16 <210> 3723 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3723 atacctcatg ttaaag 16 <210> 3724 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3724 tgatacctca tgttaa 16 <210> 3725 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3725 agtgatacct catgtt 16 <210> 3726 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3726 atagtgatac ctcatg 16 <210> 3727 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3727 gtatagtgat acctca 16 <210> 3728 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3728 aggtatagtg atacct 16 <210> 3729 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3729 taaggtatag tgatac 16 <210> 3730 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3730 aataaggtat agtgat 16 <210> 3731 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3731 gtaaacttat tgataa 16 <210> 3732 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3732 ggcatagtaa acttat 16 <210> 3733 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3733 aattttggca tagtaa 16 <210> 3734 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3734 ggcaattaat gaattt 16 <210> 3735 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3735 gtgaaaggca attaat 16 <210> 3736 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3736 agttaagtga aaggca 16 <210> 3737 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3737 cccaaaagtt aagtga 16 <210> 3738 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3738 tatggtccca aaagtt 16 <210> 3739 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3739 atttattatg gtccca 16 <210> 3740 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3740 gttatggcaa tacatt 16 <210> 3741 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3741 attaatgtta tggcaa 16 <210> 3742 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3742 gtagtttatt aatgtt 16 <210> 3743 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3743 tgtaaggtag tttatt 16 <210> 3744 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3744 tggttttgta aggtag 16 <210> 3745 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3745 aattggtggt tttgta 16 <210> 3746 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3746 gattttaatt ggtggt 16 <210> 3747 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3747 gatgtaaata acactt 16 <210> 3748 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3748 ttgacagatg taaata 16 <210> 3749 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3749 tttatgttga cagatg 16 <210> 3750 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3750 agtagattta tgttga 16 <210> 3751 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3751 cctgaatata atgaat 16 <210> 3752 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3752 ggactacctg aatata 16 <210> 3753 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3753 accatcaagc ctccca 16 <210> 3754 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3754 ccccttacca tcaagc 16 <210> 3755 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3755 tgtagtcccc ttacca 16 <210> 3756 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3756 attgaatgta gtcccc 16 <210> 3757 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3757 gattagcaag tgaatg 16 <210> 3758 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3758 tttgtagatt agcaag 16 <210> 3759 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3759 aagaggttct cagtaa 16 <210> 3760 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3760 gtccataaga ggttct 16 <210> 3761 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3761 tacctggtcc ataaga 16 <210> 3762 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3762 tcctaatacc tggtcc 16 <210> 3763 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3763 tacttttcct aatacc 16 <210> 3764 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3764 cgttactact tttcct 16 <210> 3765 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3765 ctgagactgc ttctcg 16 <210> 3766 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3766 tgaaggctga gactgc 16 <210> 3767 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3767 taaattatat gaaggc 16 <210> 3768 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3768 gtaattgttt gataat 16 <210> 3769 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3769 tactaacaaa tgtgta 16 <210> 3770 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3770 gtaatttact aacaaa 16 <210> 3771 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3771 ataagtgtaa tttact 16 <210> 3772 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3772 gttgtaataa gtgtaa 16 <210> 3773 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3773 gtgataaata taattc 16 <210> 3774 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3774 catgtaattg tgataa 16 <210> 3775 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3775 gtatatttaa gaacag 16 <210> 3776 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3776 ttgtgataag tatatt 16 <210> 3777 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3777 tggaattaaa ttgtga 16 <210> 3778 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3778 aagccgtgga attaaa 16 <210> 3779 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3779 cattgtaagc cgtgga 16 <210> 3780 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3780 tatgatcatt gtaagc 16 <210> 3781 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3781 tatagttatg atcatt 16 <210> 3782 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3782 gacataacat ttaatc 16 <210> 3783 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3783 acttatgaca taacat 16 <210> 3784 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3784 gttactactt atgaca 16 <210> 3785 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3785 gtaacagtta ctactt 16 <210> 3786 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3786 gcttatttgt aacagt 16 <210> 3787 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3787 ttcacagctt atttgt 16 <210> 3788 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3788 gttcttttca cagctt 16 <210> 3789 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3789 ggagtggttc ttttca 16 <210> 3790 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3790 atgctaggag tggttc 16 <210> 3791 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3791 tgactaatgc taggag 16 <210> 3792 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3792 atagagtgac taatgc 16 <210> 3793 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3793 gagagaatag agtgac 16 <210> 3794 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3794 attgatatgt aaaacg 16 <210> 3795 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3795 caattaattg atatgt 16 <210> 3796 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3796 ccttttaact tccaat 16 <210> 3797 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3797 cctggtcctt ttaact 16 <210> 3798 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3798 gagtttcctg gtcctt 16 <210> 3799 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3799 atgtctgagt ttcctg 16 <210> 3800 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3800 tactgtatgt ctgagt 16 <210> 3801 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3801 ccatacattc tatata 16 <210> 3802 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3802 tataagccat acattc 16 <210> 3803 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3803 attcattata agccat 16 <210> 3804 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3804 cattgagtta actaat 16 <210> 3805 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3805 aatttgcatt gagtta 16 <210> 3806 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3806 tgaagttact tctggg 16 <210> 3807 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3807 agtgaagtta cttctg 16 <210> 3808 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3808 taagtgaagt tacttc 16 <210> 3809 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3809 gttttaagtg aagtta 16 <210> 3810 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3810 aagttttaag tgaagt 16 <210> 3811 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3811 gtttttctac aaaagt 16 <210> 3812 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3812 atgctattat cttgtt 16 <210> 3813 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3813 tgatgctatt atcttg 16 <210> 3814 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3814 tttgatgcta ttatct 16 <210> 3815 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3815 gtctttgatg ctatta 16 <210> 3816 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3816 aggtctttga tgctat 16 <210> 3817 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3817 gaaggtcttt gatgct 16 <210> 3818 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3818 gagaaggtct ttgatg 16 <210> 3819 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3819 tggagaaggt ctttga 16 <210> 3820 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3820 tctggagaag gtcttt 16 <210> 3821 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3821 ggtctggaga aggtct 16 <210> 3822 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3822 acggtctgga gaaggt 16 <210> 3823 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3823 ccacggtctg gagaag 16 <210> 3824 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3824 ttccacggtc tggaga 16 <210> 3825 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3825 tcttccacgg tctgga 16 <210> 3826 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3826 ggtcttccac ggtctg 16 <210> 3827 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3827 ttggtcttcc acggtc 16 <210> 3828 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3828 tattggtctt ccacgg 16 <210> 3829 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3829 tatattggtc ttccac 16 <210> 3830 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3830 tttatattgg tcttcc 16 <210> 3831 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3831 tgtttatatt ggtctt 16 <210> 3832 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3832 attgtttata ttggtc 16 <210> 3833 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3833 taattgttta tattgg 16 <210> 3834 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3834 ggtttaattg tttata 16 <210> 3835 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3835 gttggtttaa ttgttt 16 <210> 3836 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3836 ctgttggttt aattgt 16 <210> 3837 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3837 tgctgttggt ttaatt 16 <210> 3838 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3838 tatgctgttg gtttaa 16 <210> 3839 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3839 actatgctgt tggttt 16 <210> 3840 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3840 tgactatgct gttggt 16 <210> 3841 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3841 tttgactatg ctgttg 16 <210> 3842 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3842 tatttgacta tgctgt 16 <210> 3843 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3843 tttatttgac tatgct 16 <210> 3844 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3844 cttttatttg actatg 16 <210> 3845 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3845 gagctgattt tctatt 16 <210> 3846 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3846 ctgagctgat tttcta 16 <210> 3847 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3847 ttctgagctg attttc 16 <210> 3848 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3848 ccttctgagc tgattt 16 <210> 3849 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3849 gtccttctga gctgat 16 <210> 3850 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3850 tagtccttct gagctg 16 <210> 3851 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3851 actagtcctt ctgagc 16 <210> 3852 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3852 atactagtcc ttctga 16 <210> 3853 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3853 gaatactagt ccttct 16 <210> 3854 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3854 ttgaatacta gtcctt 16 <210> 3855 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3855 tcttgaatac tagtcc 16 <210> 3856 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3856 gttcttgaat actagt 16 <210> 3857 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3857 gggttcttga atacta 16 <210> 3858 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3858 gtgggttctt gaatac 16 <210> 3859 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3859 ctgtgggttc ttgaat 16 <210> 3860 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3860 ttctgtgggt tcttga 16 <210> 3861 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3861 aaatttctgt gggttc 16 <210> 3862 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3862 agaaatttct gtgggt 16 <210> 3863 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3863 tagagaaatt tctgtg 16 <210> 3864 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3864 gatagagaaa tttctg 16 <210> 3865 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3865 gcttggaaga tagaga 16 <210> 3866 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3866 tggcttggaa gataga 16 <210> 3867 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3867 cttggcttgg aagata 16 <210> 3868 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3868 ctcttggctt ggaaga 16 <210> 3869 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3869 tgctcttggc ttggaa 16 <210> 3870 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3870 ttggtgctct tggctt 16 <210> 3871 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3871 tcttggtgct cttggc 16 <210> 3872 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3872 gttcttggtg ctcttg 16 <210> 3873 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3873 tagttcttgg tgctct 16 <210> 3874 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3874 agtagttctt ggtgct 16 <210> 3875 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3875 ggagtagttc ttggtg 16 <210> 3876 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3876 agggagtagt tcttgg 16 <210> 3877 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3877 aaagggagta gttctt 16 <210> 3878 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3878 agaaagggag tagttc 16 <210> 3879 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3879 gaagaaaggg agtagt 16 <210> 3880 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3880 ctgaagaaag ggagta 16 <210> 3881 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3881 tcaactgaag aaaggg 16 <210> 3882 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3882 tcattgaagt tttgtg 16 <210> 3883 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3883 cgtttcattg aagttt 16 <210> 3884 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3884 ttgtagttct cccacg 16 <210> 3885 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3885 atttgtagtt ctccca 16 <210> 3886 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3886 atatttgtag ttctcc 16 <210> 3887 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3887 ccatatttgt agttct 16 <210> 3888 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3888 aaccatattt gtagtt 16 <210> 3889 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3889 ccaaaaccat atttgt 16 <210> 3890 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3890 ctcccaaaac catatt 16 <210> 3891 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3891 gcctcccaaa accata 16 <210> 3892 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3892 aagcctccca aaacca 16 <210> 3893 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3893 tcaagcctcc caaaac 16 <210> 3894 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3894 tccatcaagc ctccca 16 <210> 3895 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3895 tctccatcaa gcctcc 16 <210> 3896 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3896 attctccatc aagcct 16 <210> 3897 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3897 aaattctcca tcaagc 16 <210> 3898 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3898 accaaaattc tccatc 16 <210> 3899 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3899 caaccaaaat tctcca 16 <210> 3900 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3900 cccaaccaaa attctc 16 <210> 3901 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3901 ggcccaacca aaattc 16 <210> 3902 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3902 taggcccaac caaaat 16 <210> 3903 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3903 tctaggccca accaaa 16 <210> 3904 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3904 tctctaggcc caacca 16 <210> 3905 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3905 cttctctagg cccaac 16 <210> 3906 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3906 atatcttctc taggcc 16 <210> 3907 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3907 gtatatcttc tctagg 16 <210> 3908 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3908 gagtatatct tctcta 16 <210> 3909 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3909 tggagtatat cttctc 16 <210> 3910 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3910 tatggagtat atcttc 16 <210> 3911 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3911 actatggagt atatct 16 <210> 3912 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3912 tcactatgga gtatat 16 <210> 3913 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3913 cttcactatg gagtat 16 <210> 3914 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3914 tgcttcacta tggagt 16 <210> 3915 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3915 attgcttcac tatgga 16 <210> 3916 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3916 agattgcttc actatg 16 <210> 3917 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3917 ttagattgct tcacta 16 <210> 3918 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3918 aatttagattg cttcac 16 <210> 3919 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3919 ataattagat tgcttc 16 <210> 3920 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3920 acataattag attgct 16 <210> 3921 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3921 cgtaaaacat aattag 16 <210> 3922 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3922 cttccaactc aattcg 16 <210> 3923 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3923 gtcttccaac tcaatt 16 <210> 3924 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3924 cagtcttcca actcaa 16 <210> 3925 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3925 tccagtcttc caactc 16 <210> 3926 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3926 tttccagtct tccaac 16 <210> 3927 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3927 gtctttccag tcttcc 16 <210> 3928 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3928 ttgttgtctt tccagt 16 <210> 3929 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3929 ataatgtttg ttgtct 16 <210> 3930 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3930 cgtatagttg gtttcg 16 <210> 3931 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3931 gcaactagat gtagcg 16 <210> 3932 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3932 tcgcaactag atgtag 16 <210> 3933 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3933 aatcgcaact agatgt 16 <210> 3934 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3934 gtaatcgcaa ctagat 16 <210> 3935 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3935 cagtaatcgc aactag 16 <210> 3936 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3936 gccagtaatc gcaact 16 <210> 3937 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3937 ttgccagtaa tcgcaa 16 <210> 3938 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3938 cattgccagt aatcgc 16 <210> 3939 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3939 gacattgcca gtaatc 16 <210> 3940 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3940 gggacattgc cagtaa 16 <210> 3941 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3941 tccgggattg cattgg 16 <210> 3942 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3942 tttccgggat tgcatt 16 <210> 3943 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3943 gttttccggg attgca 16 <210> 3944 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3944 ttgttttccg ggattg 16 <210> 3945 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3945 ctttgttttc cgggat 16 <210> 3946 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3946 atctttgttt tccggg 16 <210> 3947 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3947 aaatctttgt tttccg 16 <210> 3948 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3948 acaccaaatc tttgtt 16 <210> 3949 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3949 aaacaccaaa tctttg 16 <210> 3950 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3950 caagtagaaa acacca 16 <210> 3951 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3951 cccaagtaga aaacac 16 <210> 3952 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3952 atcccaagta gaaaac 16 <210> 3953 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3953 tgatcccaag tagaaa 16 <210> 3954 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3954 tgtgatccca agtaga 16 <210> 3955 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3955 ctgatcaaat atgttg 16 <210> 3956 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3956 gactgatcaa atatgt 16 <210> 3957 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3957 aagactgatc aaatat 16 <210> 3958 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3958 cataaaaaga ctgatc 16 <210> 3959 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3959 gatcataaaa agactg 16 <210> 3960 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3960 tagatcataa aaagac 16 <210> 3961 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3961 gatagatcat aaaaag 16 <210> 3962 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3962 gcgatagatc ataaaa 16 <210> 3963 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3963 cagcgataga tcataa 16 <210> 3964 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3964 tttgcagcga tagatc 16 <210> 3965 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3965 ggtttgcagc gataga 16 <210> 3966 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3966 cactggtttg cagcga 16 <210> 3967 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3967 ttcactggtt tgcagc 16 <210> 3968 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3968 atttcactgg tttgca 16 <210> 3969 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3969 tgatttcact ggtttg 16 <210> 3970 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3970 tagttcttct cagttc 16 <210> 3971 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3971 tgtagttctt ctcagt 16 <210> 3972 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3972 tatgtagttc ttctca 16 <210> 3973 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3973 tatatgtagt tcttct 16 <210> 3974 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3974 gtttatatgt agttct 16 <210> 3975 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3975 gtagtttata tgtagt 16 <210> 3976 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3976 cttgtagttt atatgt 16 <210> 3977 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3977 gacttgtagt ttatat 16 <210> 3978 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3978 ttgacttgta gtttat 16 <210> 3979 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3979 tttttgactt gtagtt 16 <210> 3980 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3980 catttttgac ttgtag 16 <210> 3981 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3981 cctcttcatt tttgac 16 <210> 3982 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3982 gacatattct ttacct 16 <210> 3983 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3983 gtgacatatt ctttac 16 <210> 3984 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3984 ttcaagtgac atattc 16 <210> 3985 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3985 agttcaagtg acatat 16 <210> 3986 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3986 tgagttcaag tgacat 16 <210> 3987 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3987 gttgagttca agtgac 16 <210> 3988 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3988 gagttgagtt caagtg 16 <210> 3989 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3989 ttgagttgag ttcaag 16 <210> 3990 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3990 ttttgagttg agttca 16 <210> 3991 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3991 agttttgagt tgagtt 16 <210> 3992 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3992 caagttttga gttgag 16 <210> 3993 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3993 ttcaagtttt gagttg 16 <210> 3994 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3994 ctttcaagtt ttgagt 16 <210> 3995 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3995 ggctttcaag ttttga 16 <210> 3996 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3996 gaggctttca agtttt 16 <210> 3997 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3997 aggagcttt caagtt 16 <210> 3998 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3998 ctaggaggct ttcaag 16 <210> 3999 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 3999 ttctaggagg ctttca 16 <210> 4000 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4000 tcttctagga ggcttt 16 <210> 4001 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4001 tttcttctag gaggct 16 <210> 4002 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4002 gttgaagtag aatttt 16 <210> 4003 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4003 gttgctcttc taaata 16 <210> 4004 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4004 tagttgctct tctaaa 16 <210> 4005 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4005 gttagttgct cttcta 16 <210> 4006 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4006 tagttagttg ctcttc 16 <210> 4007 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4007 aagttagtta gttgct 16 <210> 4008 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4008 ttaagttagt tagttg 16 <210> 4009 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4009 aattaagtta gttagt 16 <210> 4010 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4010 tgaattaagt tagtta 16 <210> 4011 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4011 tttgaattaa gttagt 16 <210> 4012 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4012 ggttgatttt gaatta 16 <210> 4013 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4013 caggttgatt ttgaat 16 <210> 4014 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4014 ttcaggttga ttttga 16 <210> 4015 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4015 gtttcaggtt gatttt 16 <210> 4016 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4016 gagtttcagg ttgatt 16 <210> 4017 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4017 tggagtttca ggttga 16 <210> 4018 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4018 ttctggagtt tcaggt 16 <210> 4019 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4019 tgttctggag tttcag 16 <210> 4020 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4020 ggtgttctgg agtttc 16 <210> 4021 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4021 tgggtgttct ggagtt 16 <210> 4022 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4022 tctgggtgtt ctggag 16 <210> 4023 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4023 cttctgggtg ttctgg 16 <210> 4024 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4024 tacttctggg tgttct 16 <210> 4025 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4025 gttacttctg ggtgtt 16 <210> 4026 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4026 aagttacttc tgggtg 16 <210> 4027 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4027 ccatcatgtt ttacat 16 <210> 4028 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4028 tgccatcatg ttttac 16 <210> 4029 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4029 gaatgccatc atgttt 16 <210> 4030 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4030 aggaatgcca tcatgt 16 <210> 4031 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4031 gcaggaatgc catcat 16 <210> 4032 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4032 cagcaggaat gccatc 16 <210> 4033 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4033 ttcagcagga atgcca 16 <210> 4034 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4034 cattcagcag gaatgc 16 <210> 4035 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4035 tacattcagc aggaat 16 <210> 4036 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4036 ggtacattca gcagga 16 <210> 4037 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4037 gtggtacatt cagcag 16 <210> 4038 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4038 tggtggtaca ttcagc 16 <210> 4039 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4039 tataaatggt ggtaca 16 <210> 4040 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4040 gttataaatg gtggta 16 <210> 4041 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4041 ctgtttataaa tggtgg 16 <210> 4042 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4042 ctctgttata aatggt 16 <210> 4043 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4043 acctctgtta taaatg 16 <210> 4044 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4044 tcacctctgt tataaa 16 <210> 4045 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4045 gttcacctct gttata 16 <210> 4046 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4046 atgttcacct ctgtta 16 <210> 4047 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4047 ttgtatgttc acctct 16 <210> 4048 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4048 acttgtatgt tcacct 16 <210> 4049 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4049 ccacttgtat gttcac 16 <210> 4050 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4050 tgccacttgt atgttc 16 <210> 4051 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4051 catgccactt gtatgt 16 <210> 4052 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4052 tacatgccac ttgtat 16 <210> 4053 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4053 catcatgcc acttgt 16 <210> 4054 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4054 ggcatacatg ccactt 16 <210> 4055 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4055 atggcataca tgccac 16 <210> 4056 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4056 tgatggcata catgcc 16 <210> 4057 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4057 tctgatggca tacatg 16 <210> 4058 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4058 ggtctgatgg cataca 16 <210> 4059 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4059 tgggtctgat ggcata 16 <210> 4060 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4060 gttgctgggt ctgatg 16 <210> 4061 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4061 gagagttgct gggtct 16 <210> 4062 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4062 ttgagagttg ctgggt 16 <210> 4063 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4063 acttgagagt tgctgg 16 <210> 4064 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4064 aaaaacttga gagttg 16 <210> 4065 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4065 tgaaaaactt gagagt 16 <210> 4066 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4066 catgaaaaac ttgaga 16 <210> 4067 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4067 gacatgaaaa acttga 16 <210> 4068 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4068 tcacagtaga catgaa 16 <210> 4069 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4069 catcacagta gacatg 16 <210> 4070 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4070 aacatcacag tagaca 16 <210> 4071 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4071 ataacatcac agtaga 16 <210> 4072 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4072 atataacatc acagta 16 <210> 4073 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4073 tgatataaca tcacag 16 <210> 4074 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4074 cctgatataa catcac 16 <210> 4075 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4075 tacctgatat aacatc 16 <210> 4076 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4076 actacctgat ataaca 16 <210> 4077 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4077 ggactacctg atataa 16 <210> 4078 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4078 atggactacc tgatat 16 <210> 4079 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4079 ccatggacta cctgat 16 <210> 4080 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4080 atgtccatgg actacc 16 <210> 4081 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4081 taatgtccat ggacta 16 <210> 4082 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4082 attaatgtcc atggac 16 <210> 4083 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4083 gaattaatgt ccatgg 16 <210> 4084 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4084 ttgaattaat gtccat 16 <210> 4085 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4085 tgttgaatta atgtcc 16 <210> 4086 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4086 gatgttgaat taatgt 16 <210> 4087 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4087 tcgatgttga attaat 16 <210> 4088 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4088 attcgatgtt gaatta 16 <210> 4089 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4089 ctattcgatg ttgaat 16 <210> 4090 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4090 atctattcga tgttga 16 <210> 4091 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4091 atccatctat tcgatg 16 <210> 4092 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4092 tgatccatct attcga 16 <210> 4093 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4093 tgtgatccat ctattc 16 <210> 4094 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4094 tttgtgatcc atctat 16 <210> 4095 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4095 gttttgtgat ccatct 16 <210> 4096 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4096 aagttttgtg atccat 16 <210> 4097 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4097 tgaagttttg tgatcc 16 <210> 4098 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4098 attgaagttt tgtgat 16 <210> 4099 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4099 caacattttg gttgat 16 <210> 4100 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4100 gatcaacatt ttggtt 16 <210> 4101 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4101 gatggatcaa catttt 16 <210> 4102 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4102 gttggatgga tcaaca 16 <210> 4103 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4103 ctgttggatg gatcaa 16 <210> 4104 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4104 atctgttgga tggatc 16 <210> 4105 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4105 ctgaatctgt tggatg 16 <210> 4106 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4106 caaagctttc tgaatc 16 <210> 4107 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4107 ctcagttcat tcaaag 16 <210> 4108 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4108 gcctcagttc attcaa 16 <210> 4109 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4109 ttgcctcagt tcattc 16 <210> 4110 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4110 atttgcctca gttcat 16 <210> 4111 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4111 aaatttgcct cagttc 16 <210> 4112 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4112 tttaaatttg cctcag 16 <210> 4113 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4113 cttttaaatt tgcctc 16 <210> 4114 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4114 gccttttaaa tttgcc 16 <210> 4115 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4115 gtttaaatta ttgcct 16 <210> 4116 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4116 atgaggttaa tgttta 16 <210> 4117 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4117 gaatgaggtt aatgtt 16 <210> 4118 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4118 tggaatgagg ttaatg 16 <210> 4119 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4119 cttggaatga ggttaa 16 <210> 4120 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4120 aacttggaat gaggtt 16 <210> 4121 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4121 cattaacttg gaatga 16 <210> 4122 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4122 cacattaact tggaat 16 <210> 4123 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4123 accacattaa cttgga 16 <210> 4124 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4124 ttagaccaca ttaact 16 <210> 4125 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4125 tattagacca cattaa 16 <210> 4126 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4126 attattagac cacat 16 <210> 4127 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4127 taccagatta ttagac 16 <210> 4128 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4128 aataccagat tattag 16 <210> 4129 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4129 ttaataccag attatt 16 <210> 4130 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4130 atttaatacc agatta 16 <210> 4131 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4131 ggattataata ccagat 16 <210> 4132 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4132 aaggatttaa taccag 16 <210> 4133 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4133 ttaaggattt aatacc 16 <210> 4134 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4134 ctcttaagga tttaat 16 <210> 4135 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4135 ttctcttaag gatta 16 <210> 4136 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4136 gctttctctt aaggat 16 <210> 4137 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4137 aagctttctc ttaagg 16 <210> 4138 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4138 tcaagctttc tcttaa 16 <210> 4139 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4139 agtgacttta agataa 16 <210> 4140 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4140 acagtgactt taagat 16 <210> 4141 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4141 agacagtgac tttaag 16 <210> 4142 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4142 aaataagacag tgactt 16 <210> 4143 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4143 ttaaatagac agtgac 16 <210> 4144 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4144 tcttaaatag acagtg 16 <210> 4145 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4145 aatcttaaat agacag 16 <210> 4146 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4146 ttaatcttaa atagac 16 <210> 4147 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4147 gtttaatctt aaatag 16 <210> 4148 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4148 gtatgtttaa tcttaa 16 <210> 4149 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4149 attgtatgtt taatct 16 <210> 4150 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4150 gtgattgtat gtttaa 16 <210> 4151 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4151 tatgtgattg tatgtt 16 <210> 4152 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4152 gttatgtgat tgtatg 16 <210> 4153 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4153 aggttatgtg attgta 16 <210> 4154 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4154 taaggttatg tgattg 16 <210> 4155 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4155 tttaaggtta tgtgat 16 <210> 4156 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4156 tctttaaggt tatgtg 16 <210> 4157 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4157 attctttaag gttatg 16 <210> 4158 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4158 gtattcttta aggtta 16 <210> 4159 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4159 cggtattctt taaggt 16 <210> 4160 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4160 aacggtattc tttaag 16 <210> 4161 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4161 taaacggtat tcttta 16 <210> 4162 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4162 tgtaaacggt attctt 16 <210> 4163 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4163 aatgtaaacg gtattc 16 <210> 4164 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4164 tctggaagca gaccta 16 <210> 4165 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4165 cttctggaag cagacc 16 <210> 4166 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4166 aaataaggta tagtga 16 <210> 4167 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4167 tagtattaag tgttaa 16 <210> 4168 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4168 tcatagtatt aagtgt 16 <210> 4169 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4169 agattccttt acaatt 16 <210> 4170 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4170 acaagattcc tttaca 16 <210> 4171 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4171 ctgacaagat tccttt 16 <210> 4172 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4172 tgtaatctga caagat 16 <210> 4173 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4173 tactgtaatc tgacaa 16 <210> 4174 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4174 tcttactgta atctga 16 <210> 4175 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4175 cattcttact gtaatc 16 <210> 4176 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4176 gttcattctt actgta 16 <210> 4177 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4177 atatgttcat tcttac 16 <210> 4178 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4178 gccacaaata tgttca 16 <210> 4179 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4179 gatgccacaa atatgt 16 <210> 4180 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4180 ctttaactcg atgcca 16 <210> 4181 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4181 aaactttaac tcgatg 16 <210> 4182 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4182 tataaacttt aactcg 16 <210> 4183 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4183 cacagcatat ttaggg 16 <210> 4184 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4184 tagaatcaca gcatat 16 <210> 4185 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4185 tattagaatc acagca 16 <210> 4186 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4186 aatgtattag aatcac 16 <210> 4187 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4187 acgaatgtat tagaat 16 <210> 4188 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4188 tacacgaatg tattag 16 <210> 4189 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4189 acctacacga atgtat 16 <210> 4190 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4190 aaaacctaca cgaatg 16 <210> 4191 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4191 ttgaaaacct acacga 16 <210> 4192 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4192 tacttgaaaa cctaca 16 <210> 4193 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4193 gtttatttct acttga 16 <210> 4194 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4194 gaggtttatt tctact 16 <210> 4195 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4195 tacgaggttt atttct 16 <210> 4196 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4196 tgttacgagg tttatt 16 <210> 4197 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4197 acttgttacg aggttt 16 <210> 4198 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4198 cagtaacttg ttacga 16 <210> 4199 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4199 gttcagtaac ttgtta 16 <210> 4200 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4200 tcaggctgtt taaacg 16 <210> 4201 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4201 ttgtcaggct gtttaa 16 <210> 4202 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4202 tatacatgct tgtcag 16 <210> 4203 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4203 gtctttgttt attgaa 16 <210> 4204 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4204 tgggtctttg tttatt 16 <210> 4205 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4205 gactgggtct ttgttt 16 <210> 4206 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4206 ataatttag gactgg 16 <210> 4207 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4207 tctataattt agggac 16 <210> 4208 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4208 cgataaacat gcaaga 16 <210> 4209 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4209 tgtcgataaa catgca 16 <210> 4210 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4210 tgatgtcgat aaacat 16 <210> 4211 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4211 ttgtgatgtc gataaa 16 <210> 4212 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4212 ctgttgtgat gtcgat 16 <210> 4213 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4213 gatctgttgt gatgtc 16 <210> 4214 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4214 agggatctgt tgtgat 16 <210> 4215 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4215 tttagggatc tgttgt 16 <210> 4216 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4216 ggatttaggg atctgt 16 <210> 4217 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4217 gattaggga tttagg 16 <210> 4218 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4218 tctttaggga tttagg 16 <210> 4219 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4219 taatctttag ggattt 16 <210> 4220 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4220 atctaatctt taggga 16 <210> 4221 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4221 tgtatctaat ctttag 16 <210> 4222 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4222 aaatttgtat ctaatc 16 <210> 4223 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4223 gtaaaaaatt tgtatc 16 <210> 4224 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4224 gtggtaaaaa atttgt 16 <210> 4225 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4225 gatactgtgg taaaaa 16 <210> 4226 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4226 agtgatactg tggtaa 16 <210> 4227 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4227 acaagtgata ctgtgg 16 <210> 4228 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4228 ctgacaagtg atactg 16 <210> 4229 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4229 attctgacaa gtgata 16 <210> 4230 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4230 taaattctga caagtg 16 <210> 4231 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4231 tactggcagt tttaaa 16 <210> 4232 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4232 tcttactggc agtttt 16 <210> 4233 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4233 atttcttact ggcagt 16 <210> 4234 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4234 aaaatttctt actggc 16 <210> 4235 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4235 aacaaatggg tttaat 16 <210> 4236 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4236 gaatatttgc aagtct 16 <210> 4237 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4237 tcattggtag aggaat 16 <210> 4238 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4238 atattttaaa gtctcg 16 <210> 4239 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4239 gttacattat tataga 16 <210> 4240 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4240 gtgaaatgtg ttacat 16 <210> 4241 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4241 tcaccagtga aatgtg 16 <210> 4242 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4242 catgtttcac cagtga 16 <210> 4243 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4243 acaagacatg tttcac 16 <210> 4244 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4244 catatgacaa gacatg 16 <210> 4245 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4245 ctataatgca tatgac 16 <210> 4246 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4246 tcctttctat aatgca 16 <210> 4247 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4247 tgattatcct ttctat 16 <210> 4248 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4248 taactgaaag tctgat 16 <210> 4249 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4249 gtgcacaaaa atgtta 16 <210> 4250 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4250 agctatgtgc acaaaa 16 <210> 4251 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4251 gaagatagct atgtgc 16 <210> 4252 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4252 tttattgaag atagct 16 <210> 4253 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4253 tcattttagt gtatct 16 <210> 4254 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4254 ccttgatcat tttagt 16 <210> 4255 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4255 tgaatccctt gatcat 16 <210> 4256 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4256 gttgtttagt cttgaa 16 <210> 4257 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4257 aattgagttg tttagt 16 <210> 4258 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4258 gcaactaatt gagttg 16 <210> 4259 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4259 attggtgcaa ctaatt 16 <210> 4260 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4260 gttttttatt ggtgca 16 <210> 4261 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4261 ggacactgac agtttt 16 <210> 4262 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4262 caggttggac actgac 16 <210> 4263 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4263 taagtacagg ttggac 16 <210> 4264 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4264 agttattaag tacagg 16 <210> 4265 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4265 tctgtgagtt attaag 16 <210> 4266 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4266 accaaaattc tcctga 16 <210> 4267 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4267 acctgaataa ccctct 16 <210> 4268 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4268 ggtatcagaa aaagat 16 <210> 4269 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4269 agtattggta tcagaa 16 <210> 4270 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4270 ggaagatact ttgaag 16 <210> 4271 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4271 aatgtgggaa gatact 16 <210> 4272 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4272 cagataatag ctaata 16 <210> 4273 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4273 tcattgcaga taatag 16 <210> 4274 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4274 gattcggatt tttaaa 16 <210> 4275 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4275 atttgggatt cggatt 16 <210> 4276 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4276 acgcttattt gggatt 16 <210> 4277 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4277 tctagagaga aaacgc 16 <210> 4278 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4278 agttaagagg ttttcg 16 <210> 4279 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4279 cattatagtt aagagg 16 <210> 4280 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4280 cactttcatt attagtt 16 <210> 4281 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4281 tagaatgaac actttc 16 <210> 4282 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4282 ttgaactaga atgaac 16 <210> 4283 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4283 acctgattga actaga 16 <210> 4284 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4284 taaaatacct gattga 16 <210> 4285 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4285 tagaggtaaa atacct 16 <210> 4286 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4286 gaagattaga ggtaaa 16 <210> 4287 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4287 tctgaggaag attaga 16 <210> 4288 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4288 tatacactac caaaaa 16 <210> 4289 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4289 ataatctata cactac 16 <210> 4290 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4290 taagtcccaa ttttaa 16 <210> 4291 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4291 ccagttttaa ataatc 16 <210> 4292 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4292 tgtatcccag ttttaa 16 <210> 4293 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4293 gttttagatg catgta 16 <210> 4294 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4294 tacagtgttt tagatg 16 <210> 4295 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4295 ttccccgtaa gtttat 16 <210> 4296 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4296 ctgtatttcc ccgtaa 16 <210> 4297 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4297 tagttactgt tactgt 16 <210> 4298 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4298 cgtatgtagt tactgt 16 <210> 4299 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4299 aatgggtaca gactcg 16 <210> 4300 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4300 gcaatttaat gggtac 16 <210> 4301 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4301 gatagatatg caattt 16 <210> 4302 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4302 aaaggagata gatatg 16 <210> 4303 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4303 cctcctaaag gagata 16 <210> 4304 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4304 caccagcctc ctaaag 16 <210> 4305 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4305 tttgtgatcc caagta 16 <210> 4306 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4306 gctttgtgat cccaag 16 <210> 4307 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4307 ttgctttgtg atccca 16 <210> 4308 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4308 ttttgctttg tgatcc 16 <210> 4309 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4309 ccttttgctt tgtgat 16 <210> 4310 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4310 gtccttttgc tttgtg 16 <210> 4311 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4311 gtgtcctttt gctttg 16 <210> 4312 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4312 gaagtgtcct tttgct 16 <210> 4313 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4313 ttgaagtgtc cttttg 16 <210> 4314 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4314 agttgaagtg tccttt 16 <210> 4315 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4315 acagttgaag tgtcct 16 <210> 4316 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4316 ggacagttga agtgtc 16 <210> 4317 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4317 ctggacagtt gaagtg 16 <210> 4318 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4318 ctctggacag ttgaag 16 <210> 4319 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4319 ccctctggac agttga 16 <210> 4320 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4320 aaccctctgg acagtt 16 <210> 4321 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4321 ataaccctct ggacag 16 <210> 4322 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4322 gaataaccct ctggac 16 <210> 4323 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4323 ctgaataacc ctctgg 16 <210> 4324 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4324 tcctgaataa ccctct 16 <210> 4325 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4325 cctcctgaat aaccct 16 <210> 4326 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4326 accaccagcc tcctga 16 <210> 4327 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4327 atgccaccac caggct 16 <210> 4328 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4328 tcatgccacc accagc 16 <210> 4329 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4329 catcatgcca ccacca 16 <210> 4330 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4330 cactcatcat gccacc 16 <210> 4331 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4331 tccacactca tcatgc 16 <210> 4332 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4332 tctccacact catcat 16 <210> 4333 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4333 tttctccaca ctcatc 16 <210> 4334 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4334 gttttctcca cactca 16 <210> 4335 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4335 gttgttttct ccacac 16 <210> 4336 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4336 aggttgtttt ctccac 16 <210> 4337 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4337 ttaggttgtt ttctcc 16 <210> 4338 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4338 taccatttag gttgtt 16 <210> 4339 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4339 tttaccattt aggttg 16 <210> 4340 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4340 tattaccat ttaggt 16 <210> 4341 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4341 ttgttatatt taccat 16 <210> 4342 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4342 gtttgttata tttacc 16 <210> 4343 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4343 cttggtttgt tatatt 16 <210> 4344 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4344 ctcttggttt gttata 16 <210> 4345 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4345 tgctcttggt ttgtta 16 <210> 4346 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4346 tttgctcttg gtttgt 16 <210> 4347 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4347 gattttgctc ttggtt 16 <210> 4348 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4348 cttagatttt gctctt 16 <210> 4349 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4349 ggcttagatt ttgctc 16 <210> 4350 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4350 ctggcttaga ttttgc 16 <210> 4351 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4351 ctctggctta gatttt 16 <210> 4352 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4352 ctctctggct tagatt 16 <210> 4353 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4353 tcctctctgg cttaga 16 <210> 4354 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4354 tctcctctct ggctta 16 <210> 4355 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4355 taatcctctt ctcctc 16 <210> 4356 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4356 gataatcctc ttctcc 16 <210> 4357 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4357 aagataatcc tcttct 16 <210> 4358 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4358 ccaagataat cctctt 16 <210> 4359 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4359 ttccaagata atcctc 16 <210> 4360 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4360 acttccaaga taatcc 16 <210> 4361 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4361 agacttccaa gataat 16 <210> 4362 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4362 tgagacttcc aagata 16 <210> 4363 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4363 tttgagactt ccaaga 16 <210> 4364 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4364 attttgagac ttccaa 16 <210> 4365 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4365 ttccattttg agactt 16 <210> 4366 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4366 ccttccattt tgagac 16 <210> 4367 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4367 aaccttccat tttgag 16 <210> 4368 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4368 ataaccttcc attttg 16 <210> 4369 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4369 gtataacctt ccattt 16 <210> 4370 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4370 gagtataacc ttccat 16 <210> 4371 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4371 tatagagtat aacctt 16 <210> 4372 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4372 tttataggt ataacc 16 <210> 4373 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4373 gattttatag agtata 16 <210> 4374 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4374 ttgattttat agagta 16 <210> 4375 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4375 ggttgatttt atagag 16 <210> 4376 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4376 ttggttgatt ttatag 16 <210> 4377 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4377 taatgtttaa attattgcct 20 <210> 4378 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4378 ggttaatgtt taaattattg 20 <210> 4379 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4379 aggttaatgt ttaaattatt 20 <210> 4380 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4380 gaggttaatg tttaaattat 20 <210> 4381 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4381 tgaggttaat gtttaaatta 20 <210> 4382 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4382 aatgaggtta atgtttaaat 20 <210> 4383 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4383 gaatgaggtt aatgtttaaa 20 <210> 4384 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4384 ggaatgaggt taatgtttaa 20 <210> 4385 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4385 tggaatgagg ttaatgttta 20 <210> 4386 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4386 ttggaatgag gttaatgttt 20 <210> 4387 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4387 cttggaatga ggttaatgtt 20 <210> 4388 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4388 taacttggaa tgaggttaat 20 <210> 4389 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4389 ttaacttgga atgaggttaa 20 <210> 4390 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4390 attaacttgg aatgaggtta 20 <210> 4391 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4391 cattaacttg gaatgaggtt 20 <210> 4392 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4392 acattaactt ggaatgaggt 20 <210> 4393 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4393 accacattaa cttggaatga 20 <210> 4394 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4394 gaccacatta acttggaatg 20 <210> 4395 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4395 agaccacatt aacttggaat 20 <210> 4396 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4396 tagaccacat taacttggaa 20 <210> 4397 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4397 ttagaccaca ttaacttgga 20 <210> 4398 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4398 attagaccac attaacttgg 20 <210> 4399 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4399 tattagacca cattaacttg 20 <210> 4400 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4400 ttattagacc acattaactt 20 <210> 4401 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4401 attattagac cacattaact 20 <210> 4402 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4402 gattattaga ccacattaac 20 <210> 4403 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4403 aataccagat tattagacca 20 <210> 4404 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4404 taataccaga ttattagacc 20 <210> 4405 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4405 tttaatacca gattattaga 20 <210> 4406 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4406 atttaatacc agattattag 20 <210> 4407 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4407 gattaatac cagattatta 20 <210> 4408 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4408 taaggattta ataccagatt 20 <210> 4409 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4409 tttctcttaa ggatttaata 20 <210> 4410 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4410 ctttctctta aggatttaat 20 <210> 4411 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4411 agctttctct taaggattta 20 <210> 4412 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4412 aagctttctc ttaaggattt 20 <210> 4413 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4413 tcaagctttc tcttaaggat 20 <210> 4414 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4414 ttctcaagct ttctcttaag 20 <210> 4415 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4415 tttctcaagc tttctcttaa 20 <210> 4416 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4416 tctatttctc aagctttctc 20 <210> 4417 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4417 atctatttct caagctttct 20 <210> 4418 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4418 aatctatttc tcaagctttc 20 <210> 4419 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4419 aaatctattt ctcaagcttt 20 <210> 4420 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4420 aaaatctatt tctcaagctt 20 <210> 4421 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4421 gataaaaaaa atctatttct 20 <210> 4422 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4422 tagacagtga ctttaagata 20 <210> 4423 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4423 atagacagtg actttaagat 20 <210> 4424 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4424 aataagacagt gactttaaga 20 <210> 4425 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4425 aaataagacag tgactttaag 20 <210> 4426 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4426 taaatagaca gtgactttaa 20 <210> 4427 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4427 ttaaatagac agtgacttta 20 <210> 4428 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4428 cttaaataga cagtgacttt 20 <210> 4429 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4429 tcttaaatag acagtgactt 20 <210> 4430 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4430 atcttaaata gacagtgact 20 <210> 4431 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4431 aatcttaaat agacagtgac 20 <210> 4432 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4432 taatcttaaa tagacagtga 20 <210> 4433 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4433 ttaatcttaa atagacagtg 20 <210> 4434 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4434 tttaatctta aataagacagt 20 <210> 4435 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4435 gtttaatctt aaataagacag 20 <210> 4436 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4436 tgtttaatct taaatagaca 20 <210> 4437 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4437 atgtttaatc ttaaatatagac 20 <210> 4438 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4438 ttgtatgttt aatcttaaat 20 <210> 4439 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4439 attgtatgtt taatcttaaa 20 <210> 4440 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4440 gattgtatgt ttaatcttaa 20 <210> 4441 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4441 tgattgtatg tttaatctta 20 <210> 4442 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4442 tatgtgattg tatgtttaat 20 <210> 4443 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4443 gttatgtgat tgtatgttta 20 <210> 4444 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4444 taaggttatg tgattgtatg 20 <210> 4445 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4445 ttaaggttat gtgattgtat 20 <210> 4446 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4446 ttctttaagg ttatgtgatt 20 <210> 4447 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4447 gaaatgtaaa cggtat 16 <210> 4448 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4448 ttgagaaatg taaacg 16 <210> 4449 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4449 tgattgagaa atgtaa 16 <210> 4450 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4450 tttgattgag aaatgt 16 <210> 4451 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4451 aagaattttg attgag 16 <210> 4452 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4452 ataagaattt tgattg 16 <210> 4453 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4453 caaatagtat tataag 16 <210> 4454 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4454 cccacatcac aaaatt 16 <210> 4455 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4455 gattcccaca tcacaa 16 <210> 4456 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4456 ttgattccca catcac 16 <210> 4457 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4457 aattgattcc cacatc 16 <210> 4458 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4458 aaaattgatt cccaca 16 <210> 4459 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4459 ctaaaattga ttccca 16 <210> 4460 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4460 catctaaaat tgattc 16 <210> 4461 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4461 accatctaaa attgat 16 <210> 4462 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4462 tgaccatcta aaattg 16 <210> 4463 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4463 tgtgaccatc taaaat 16 <210> 4464 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4464 attgtgacca tctaaa 16 <210> 4465 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4465 agattgtgac catcta 16 <210> 4466 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4466 ctagattgtg accatc 16 <210> 4467 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4467 atctagattg tgacca 16 <210> 4468 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4468 taatctagat tgtgac 16 <210> 4469 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4469 tataatctag attgtg 16 <210> 4470 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4470 attataatct agattg 16 <210> 4471 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4471 tgattataat ctagat 16 <210> 4472 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4472 attgattata atctag 16 <210> 4473 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4473 ctattgatta taatct 16 <210> 4474 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4474 acctattgat tataat 16 <210> 4475 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4475 tcacctattg attata 16 <210> 4476 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4476 gttcacctat tgatta 16 <210> 4477 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4477 aagttcacct attgat 16 <210> 4478 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4478 ataagttcac ctattg 16 <210> 4479 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4479 taataagttc acctat 16 <210> 4480 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4480 tttaataagt tcacct 16 <210> 4481 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4481 tattataataa gttcac 16 <210> 4482 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4482 gttatttaat aagttc 16 <210> 4483 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4483 catatgatgc ctttta 16 <210> 4484 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4484 tattagctca tatgat 16 <210> 4485 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4485 gatattagct catatg 16 <210> 4486 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4486 gtgatattag ctcata 16 <210> 4487 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4487 agttgtgata ttagct 16 <210> 4488 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4488 aaagttgtga tattag 16 <210> 4489 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4489 ggaaagttgt gatatt 16 <210> 4490 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4490 tgggaaagtt gtgata 16 <210> 4491 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4491 aaactgggaa agttgt 16 <210> 4492 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4492 ttaaactggg aaagtt 16 <210> 4493 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4493 gttttttaaa ctggga 16 <210> 4494 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4494 tagtttttta aactgg 16 <210> 4495 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4495 gagtactagt ttttta 16 <210> 4496 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4496 aagagtacta gttttt 16 <210> 4497 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4497 acaagagtac tagttt 16 <210> 4498 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4498 taacaagagt actagt 16 <210> 4499 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4499 tttaacaaga gtacta 16 <210> 4500 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4500 gttttaacaa gagtac 16 <210> 4501 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4501 gagttttaac aagagt 16 <210> 4502 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4502 tagagtttta acaaga 16 <210> 4503 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4503 gtttagagtt ttaaca 16 <210> 4504 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4504 caagtttaga gtttta 16 <210> 4505 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4505 gtcaagttta gagttt 16 <210> 4506 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4506 tagtcaagtt tagagt 16 <210> 4507 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4507 tttagtcaag tttaga 16 <210> 4508 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4508 tatttagtca agttta 16 <210> 4509 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4509 tgtatttagt caagtt 16 <210> 4510 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4510 tctgtatttta gtcaag 16 <210> 4511 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4511 cctctgtatt tagtca 16 <210> 4512 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4512 gtcctctgta tttagt 16 <210> 4513 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4513 cagtcctctg tattta 16 <210> 4514 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4514 accagtcctc tgtatt 16 <210> 4515 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4515 ttaccagtcc tctgta 16 <210> 4516 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4516 aattaccagt cctctg 16 <210> 4517 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4517 acaattacca gtcctc 16 <210> 4518 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4518 gcaaattttc agtgtt 16 <210> 4519 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4519 cgatttgtaa ttttca 16 <210> 4520 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4520 tttaaccgat ttgtaa 16 <210> 4521 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4521 gtataattta accgat 16 <210> 4522 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4522 ctagattgta taattt 16 <210> 4523 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4523 agtgttctag attgta 16 <210> 4524 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4524 tgacatagtg ttctag 16 <210> 4525 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4525 gtgtaatgac attagtg 16 <210> 4526 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4526 acaatagtgt aatgac 16 <210> 4527 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4527 gtaatttaca attagtg 16 <210> 4528 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4528 ccttcagtaa tttaca 16 <210> 4529 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4529 tacttacctt cagtaa 16 <210> 4530 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4530 ctggagaata gtttta 16 <210> 4531 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4531 ttaaacactg gagaat 16 <210> 4532 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4532 gcccagcata ttttca 16 <210> 4533 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4533 gaaaaagccc agcata 16 <210> 4534 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4534 gattttctga acttca 16 <210> 4535 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4535 gtactatctc taaaat 16 <210> 4536 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4536 taaattgtac tatctc 16 <210> 4537 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4537 cacatatttt tgtcct 16 <210> 4538 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4538 ctttcaaata gcacat 16 <210> 4539 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4539 gtatgcttct ttcaaa 16 <210> 4540 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4540 ccccttgtat gcttct 16 <210> 4541 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4541 ttccttcccc ttgtat 16 <210> 4542 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4542 tggcaattcc ttcccc 16 <210> 4543 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4543 gaatattggc aattcc 16 <210> 4544 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4544 ctaataatgg atttga 16 <210> 4545 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4545 ctatcataat ctaaat 16 <210> 4546 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4546 gtaacactat cataat 16 <210> 4547 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4547 aatttcctgt aacact 16 <210> 4548 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4548 aagttgcttt cctctt 16 <210> 4549 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4549 ggttataagt tgcttt 16 <210> 4550 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4550 taggttggtt ataagt 16 <210> 4551 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4551 agagagtag ttggtt 16 <210> 4552 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4552 ggatatagag agtagg 16 <210> 4553 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4553 aagtctggat atagag 16 <210> 4554 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4554 ctacaaaagt ctggat 16 <210> 4555 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4555 cttacctgat tttcta 16 <210> 4556 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4556 tactgactta cctgat 16 <210> 4557 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4557 ccattaaaat actgac 16 <210> 4558 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4558 ggacataacca ttaaaa 16 <210> 4559 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4559 aagatgggac atacca 16 <210> 4560 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4560 gtgtgaaaga tgggac 16 <210> 4561 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4561 agacctgtgt gaaaga 16 <210> 4562 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4562 ttttacagac ctgtgt 16 <210> 4563 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4563 cagtgttttt acagac 16 <210> 4564 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4564 taggattcag tgtttt 16 <210> 4565 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4565 gttaaagctt gtaaat 16 <210> 4566 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4566 gatccagtta aagctt 16 <210> 4567 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4567 actcatgatc cagtta 16 <210> 4568 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4568 aattttactc atgatc 16 <210> 4569 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4569 tgtgataatt ttactc 16 <210> 4570 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4570 tgctgatgtg ataatt 16 <210> 4571 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4571 gcaattttaa cagtta 16 <210> 4572 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4572 gagcctgcaa ttttaa 16 <210> 4573 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4573 tagcttcaga gcctgc 16 <210> 4574 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4574 gtttattagc ttcaga 16 <210> 4575 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4575 caggtagttt attagc 16 <210> 4576 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4576 taaatgcagg tagttt 16 <210> 4577 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4577 atggtttaaa tgcagg 16 <210> 4578 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4578 tagagccatg gtttaa 16 <210> 4579 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4579 tcacacaaag ttttag 16 <210> 4580 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4580 gtgaagtaat ttattc 16 <210> 4581 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4581 actgagagat aaaggg 16 <210> 4582 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4582 gtatatgtga ggaaac 16 <210> 4583 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4583 tttgtagtat atgtga 16 <210> 4584 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4584 attatctttg tagtat 16 <210> 4585 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4585 ataagttctg ttatta 16 <210> 4586 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4586 aatcctataa gttctg 16 <210> 4587 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4587 ctgctatgaa ttaatt 16 <210> 4588 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4588 cattggctgc tatgaa 16 <210> 4589 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4589 agatgacatt ggctgc 16 <210> 4590 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4590 ttagtaagat gacatt 16 <210> 4591 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4591 gatctaattt gaattt 16 <210> 4592 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4592 ttgagcaaag agaaac 16 <210> 4593 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4593 gaatgttgac ctttaa 16 <210> 4594 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4594 attgttgaat gttgac 16 <210> 4595 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4595 ttaattacat tgttga 16 <210> 4596 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4596 ttgtagatta attaca 16 <210> 4597 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4597 tttacattgt agatta 16 <210> 4598 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4598 cagatgttta cattgt 16 <210> 4599 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4599 cttcaccaga tgttta 16 <210> 4600 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4600 agtgcttccc tctgtc 16 <210> 4601 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4601 taaacaagtg cttccc 16 <210> 4602 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4602 tagctttttt ctaaac 16 <210> 4603 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4603 ctgacatagc tttttt 16 <210> 4604 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4604 aatacatgga ttctga 16 <210> 4605 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4605 tattagaata catgga 16 <210> 4606 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4606 gtactgcata ttagaa 16 <210> 4607 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4607 actattgtac tgcata 16 <210> 4608 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4608 ttttaaacta ttgtac 16 <210> 4609 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4609 gagagtatta ttaata 16 <210> 4610 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4610 ctgtttgaga gtatta 16 <210> 4611 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4611 gaatagctgt ttgaga 16 <210> 4612 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4612 aatcctcttg aatagc 16 <210> 4613 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4613 tttttgaatc ctcttg 16 <210> 4614 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4614 gagtttatat tatgtt 16 <210> 4615 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4615 gtttctctga gtttat 16 <210> 4616 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4616 ttaccagttt ctctga 16 <210> 4617 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4617 gattttgttt accagt 16 <210> 4618 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4618 gttttatatc tcttga 16 <210> 4619 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4619 ttggtaataa tatttg 16 <210> 4620 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4620 tggaaattgg taataa 16 <210> 4621 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4621 gtttagtgga aattgg 16 <210> 4622 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4622 atgtttgttt agtgga 16 <210> 4623 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4623 ctaacattat gtttgt 16 <210> 4624 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4624 gcactactaa cattat 16 <210> 4625 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4625 ttagcagcac tactaa 16 <210> 4626 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4626 ttgataaaaa accttt 16 <210> 4627 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4627 caaaagtagt tgataa 16 <210> 4628 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4628 ggaaaccaaa agtagt 16 <210> 4629 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4629 gaaagtatgg aaacca 16 <210> 4630 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4630 acatcataag aaggaa 16 <210> 4631 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4631 tcatagtaaa agatat 16 <210> 4632 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4632 tcatttaatc atagta 16 <210> 4633 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4633 gcaggttcat ttaatc 16 <210> 4634 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4634 gtaacatttt gctttg 16 <210> 4635 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4635 atattactat agtaac 16 <210> 4636 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4636 caatgtatat tactat 16 <210> 4637 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4637 tagacacaat gtatat 16 <210> 4638 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4638 ggtttcttca cacat 16 <210> 4639 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4639 ctcagaaatt cattgt 16 <210> 4640 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4640 cttcttccaa ctcaga 16 <210> 4641 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4641 ctaactcttc ttccaa 16 <210> 4642 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4642 aatgatctaa ctcttc 16 <210> 4643 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4643 gaaagttaaa tgatct 16 <210> 4644 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4644 atcttaaagt tactta 16 <210> 4645 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4645 tatgtgatct taaagt 16 <210> 4646 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4646 agtaactatg tgatct 16 <210> 4647 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4647 ctactaagta actatg 16 <210> 4648 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4648 tcttttctac taagta 16 <210> 4649 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4649 tattactctt ttctac 16 <210> 4650 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4650 gctgggtatt actctt 16 <210> 4651 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4651 taaagtttgc ttgctg 16 <210> 4652 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4652 ctattgtaaa gtttgc 16 <210> 4653 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4653 aaggatctat tgtaaa 16 <210> 4654 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4654 cttatttaaa aggatc 16 <210> 4655 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4655 taggacctta tttaaa 16 <210> 4656 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4656 atttcctagg acctta 16 <210> 4657 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4657 catgaatgat atttcc 16 <210> 4658 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4658 tgctggcatg aatgat 16 <210> 4659 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4659 ttttgatgct ggcatg 16 <210> 4660 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4660 ttagtttttt gatgct 16 <210> 4661 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4661 gcattattag tgttag 16 <210> 4662 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4662 tatcttgcat tattag 16 <210> 4663 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4663 atataatatc ttgcat 16 <210> 4664 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4664 cattgacagt aagaaa 16 <210> 4665 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4665 agtttttctc attgac 16 <210> 4666 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4666 atggatatct cttaac 16 <210> 4667 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4667 tatttgatgg atatct 16 <210> 4668 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4668 acattgtatt tgatgg 16 <210> 4669 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4669 gttgatacat tgtatt 16 <210> 4670 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4670 gtttaggttg atacat 16 <210> 4671 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4671 catccagttt aggttg 16 <210> 4672 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4672 ccccagcatc cagttt 16 <210> 4673 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4673 aaagaacccc agcatc 16 <210> 4674 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4674 gtgtaaaaag aacccc 16 <210> 4675 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4675 tatagggtgt aaaaag 16 <210> 4676 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4676 aggtatgtct tttata 16 <210> 4677 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4677 agtccatatt tgtatt 16 <210> 4678 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4678 taatcaagtc catatt 16 <210> 4679 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4679 atctaataat caagtc 16 <210> 4680 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4680 ccttctatat tatcta 16 <210> 4681 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4681 taataaacct tctata 16 <210> 4682 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4682 gatcacatct aagaaa 16 <210> 4683 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4683 taccatgatc acatct 16 <210> 4684 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4684 ctgcaatacc atgatc 16 <210> 4685 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4685 gttctccttt aaaact 16 <210> 4686 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4686 gagatgttc tccttt 16 <210> 4687 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4687 aaacaggaga ttgttc 16 <210> 4688 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4688 tctcttaaac aggaga 16 <210> 4689 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4689 catgtatctc ttaaac 16 <210> 4690 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4690 cgtaaatatt tcagca 16 <210> 4691 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4691 taactccgta aatatt 16 <210> 4692 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4692 gacctttaac tccgta 16 <210> 4693 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4693 tccagtgacc tttaac 16 <210> 4694 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4694 caccagtctg gagtcc 16 <210> 4695 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4695 ttctatcacc agtctg 16 <210> 4696 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4696 atcttaccaa actatt 16 <210> 4697 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4697 agaatcatct taccaa 16 <210> 4698 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4698 gaatgtaaga atcatc 16 <210> 4699 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4699 gtgttattta agaatg 16 <210> 4700 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4700 gtttatgtta aagcat 16 <210> 4701 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4701 agtaatgttt atgtta 16 <210> 4702 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4702 gtagcatttt ttcagt 16 <210> 4703 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4703 gcaaatgtag catttt 16 <210> 4704 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4704 gttgtggcaa atgtag 16 <210> 4705 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4705 tatgaagttg tggcaa 16 <210> 4706 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4706 gatttcactt gacat 16 <210> 4707 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4707 gcttgagatt tcactt 16 <210> 4708 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4708 tttggagctt gagatt 16 <210> 4709 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4709 aatatctttg gagctt 16 <210> 4710 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4710 aggaataata tctttg 16 <210> 4711 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4711 atttagtaat aggaat 16 <210> 4712 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4712 catcagattt agtaat 16 <210> 4713 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4713 gttattacat cagatt 16 <210> 4714 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4714 gcctagaatc aataaa 16 <210> 4715 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4715 aggaatgcct agaatc 16 <210> 4716 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4716 ttcagcagga atgcct 16 <210> 4717 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4717 ttacctgata taacat 16 <210> 4718 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4718 caggttttac ctgata 16 <210> 4719 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4719 cttagacagg ttttac 16 <210> 4720 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4720 attctcctta gacagg 16 <210> 4721 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4721 ctgtctattc tcctta 16 <210> 4722 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4722 taactactgt ctattc 16 <210> 4723 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4723 ttgaactaac tactgt 16 <210> 4724 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4724 agtaagttga actaac 16 <210> 4725 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4725 gtaatgagta agttga 16 <210> 4726 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4726 taatcttcct aatacg 16 <210> 4727 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4727 accaggttaa tcttcc 16 <210> 4728 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4728 atgataacca ggttaa 16 <210> 4729 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4729 cgaatactca tatata 16 <210> 4730 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4730 tttatacgaa tactca 16 <210> 4731 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4731 attatattta tacgaa 16 <210> 4732 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4732 ggtaaaagta ttatat 16 <210> 4733 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4733 gagaatattg agtaaa 16 <210> 4734 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4734 cagattattt tagagg 16 <210> 4735 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4735 tcacttcaga ttattt 16 <210> 4736 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4736 taatagtcac ttcaga 16 <210> 4737 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4737 tattgataat agtcac 16 <210> 4738 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4738 tactatttgt aatcaa 16 <210> 4739 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4739 cttgcttatt tacta 16 <210> 4740 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4740 catctgttat tttatc 16 <210> 4741 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4741 atgtgctttt tggatt 16 <210> 4742 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4742 ggatttttgt atgtgc 16 <210> 4743 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4743 catcattcat ggattt 16 <210> 4744 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4744 cttagacatc attcat 16 <210> 4745 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4745 tgagtactta gacatc 16 <210> 4746 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4746 tataagtgag tactta 16 <210> 4747 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4747 ctactttata agtgag 16 <210> 4748 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4748 tgaatgtctt ctactt 16 <210> 4749 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4749 tataataatg aatgtc 16 <210> 4750 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4750 gtactgagca tttaaa 16 <210> 4751 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4751 caaatagtac tgagca 16 <210> 4752 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4752 aatggtcaaa tagtac 16 <210> 4753 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4753 gtagtttgaa tacaaa 16 <210> 4754 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4754 tcactggtag tttgaa 16 <210> 4755 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4755 taggtagggc tttcac 16 <210> 4756 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4756 accttctag tagggc 16 <210> 4757 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4757 gagtatacct tctagg 16 <210> 4758 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4758 atcactgagt atacct 16 <210> 4759 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4759 aaacttatca ctgagt 16 <210> 4760 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4760 gctacaaaac ttatca 16 <210> 4761 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4761 tttggagcta caaaac 16 <210> 4762 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4762 agaagattg gagcta 16 <210> 4763 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4763 actattagaa gatttg 16 <210> 4764 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4764 acactcacta ttagaa 16 <210> 4765 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4765 agccttttat tttggg 16 <210> 4766 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4766 cctgtcagcc ttttat 16 <210> 4767 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4767 gacttacctg tcagcc 16 <210> 4768 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4768 attctcgact tacctg 16 <210> 4769 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4769 gtgagtattc tcgact 16 <210> 4770 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4770 aattaagtga gtattc 16 <210> 4771 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4771 taccagaatt aagtga 16 <210> 4772 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4772 gctttcttac cagaat 16 <210> 4773 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4773 tgggttgctt tcttac 16 <210> 4774 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4774 tacaagtaca aatggg 16 <210> 4775 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4775 ggtaaataca agtaca 16 <210> 4776 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4776 attgctggta aataca 16 <210> 4777 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4777 ttaaggattg ctggta 16 <210> 4778 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4778 gcttcatttt aaggat 16 <210> 4779 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4779 gtaggaagct tcattt 16 <210> 4780 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4780 gagttagtag gaagct 16 <210> 4781 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4781 gctattgagt tagtag 16 <210> 4782 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4782 cttattgcta ttgagt 16 <210> 4783 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4783 tattgtctta ttgcta 16 <210> 4784 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4784 attcactatt gtctta 16 <210> 4785 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4785 atcacaatcc ttttta 16 <210> 4786 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4786 ttcttcatca caatcc 16 <210> 4787 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4787 agattgttct tcatca 16 <210> 4788 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4788 tataaataga ttgttc 16 <210> 4789 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4789 ggttcttaat aacttt 16 <210> 4790 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4790 aagcatggtt cttaat 16 <210> 4791 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4791 ctttgtagaa aaagac 16 <210> 4792 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4792 tatgctttct ttgtag 16 <210> 4793 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4793 cttaatgtat gctttc 16 <210> 4794 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4794 gtatttgctt aatgta 16 <210> 4795 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4795 cctttggtat ttgctt 16 <210> 4796 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4796 acctggcctt tggtat 16 <210> 4797 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4797 atgtaaacct ggcctt 16 <210> 4798 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4798 cttcaaatgt aaacct 16 <210> 4799 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4799 gtaataataa tgtcac 16 <210> 4800 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4800 agacttgagt aataat 16 <210> 4801 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4801 tcctagagac ttgagt 16 <210> 4802 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4802 aagtattcct agagac 16 <210> 4803 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4803 tgtgttaagt attcct 16 <210> 4804 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4804 aagagatgtg ttaagt 16 <210> 4805 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4805 ccatatacag tcaaga 16 <210> 4806 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4806 taacatccat atacag 16 <210> 4807 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4807 ctattttatta acatcc 16 <210> 4808 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4808 tgtcagctat ttatta 16 <210> 4809 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4809 ctttactgtc agctat 16 <210> 4810 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4810 gataaacttt actgtc 16 <210> 4811 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4811 ctttatatgg ataaac 16 <210> 4812 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4812 gcaagtcttt atatgg 16 <210> 4813 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4813 ctgggtatta ctcttttcta 20 <210> 4814 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4814 cttgctgggt attactcttt 20 <210> 4815 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4815 tgcttgctgg gtattactct 20 <210> 4816 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4816 catgaatgat atttcctagg 20 <210> 4817 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4817 ggcatgaatg atatttccta 20 <210> 4818 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4818 ctggcatgaa tgatatttcc 20 <210> 4819 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4819 tgctggcatg aatgatattt 20 <210> 4820 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4820 aagtccatat ttgtatttct 20 <210> 4821 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4821 gcaaatgtag cattttttca 20 <210> 4822 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4822 ggcaaatgta gcattttttc 20 <210> 4823 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4823 ctggtccttt taacttccaa 20 <210> 4824 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4824 cctggtcctt ttaacttcca 20 <210> 4825 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4825 ttcctggtcc ttttaacttc 20 <210> 4826 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4826 tgcttaatgt atgctttctt 20 <210> 4827 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4827 ccgtaagttt atcttccttt 20 <210> 4828 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4828 ccccgtaagt ttatcttcct 20 <210> 4829 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4829 cacaaatatg ttcattctta 20 <210> 4830 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4830 aaactttaac tcgatgccac 20 <210> 4831 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4831 ataaacttta actcgatgcc 20 <210> 4832 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4832 atgcttgtca ggctgtttaa 20 <210> 4833 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4833 gtcaccatat aacttgggca 20 <210> 4834 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4834 aggtcaccat ataacttggg 20 <210> 4835 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4835 gctgggtatt actcttt 17 <210> 4836 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4836 gcatgaatga tatttcc 17 <210> 4837 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4837 ggcaaatgta gcatttt 17 <210> 4838 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4838 ctggtccttt taacttc 17 <210> 4839 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4839 gtaagtttat cttcctt 17 <210> 4840 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4840 actttaactc gatgcca 17 <210> 4841 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4841 aactttaact cgatgcc 17 <210> 4842 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4842 aaactttaac tcgatgc 17 <210> 4843 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4843 caccatataa cttgggc 17 <210> 4844 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4844 tcaccatata acttggg 17 <210> 4845 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4845 gtcaccatat aacttgg 17 <210> 4846 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4846 cttgattttg gctctggaga 20 <210> 4847 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4847 tgattttggc tctggag 17 <210> 4848 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4848 actggtttgc agcgata 17 <210> 4849 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4849 tttcactggt ttgcagcgat 20 <210> 4850 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4850 cactggtttg cagcgat 17 <210> 4851 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4851 tcactggttt gcagcga 17 <210> 4852 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4852 ttcttggtgc tcttggc 17 <210> 4853 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4853 gttcttggtg ctcttgg 17 <210> 4854 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4854 ttgtctttcc agtcttccaa 20 <210> 4855 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4855 tgttgtcttt ccagtcttcc 20 <210> 4856 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4856 cattgccagt aatcgca 17 <210> 4857 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4857 acattgccag taatcgc 17 <210> 4858 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4858 gacattgcca gtaatcg 17 <210> 4859 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4859 ctttgtgatc ccaagtagaa 20 <210> 4860 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4860 ttgtgatccc aagtaga 17 <210> 4861 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4861 tgctttgtga tcccaagtag 20 <210> 4862 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4862 tttgtgatcc caagtag 17 <210> 4863 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4863 ctttgtgatc ccaagta 17 <210> 4864 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4864 cacactcatc atgccac 17 <210> 4865 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4865 gttgttttct ccacactcat 20 <210> 4866 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4866 agattttgct cttggtt 17 <210> 4867 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4867 tagattttgc tcttggt 17 <210> 4868 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4868 ttagattttg ctcttgg 17 <210> 4869 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4869 cttagatttt gctcttg 17 <210> 4870 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4870 agattattag accacat 17 <210> 4871 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4871 cagattatta gaccaca 17 <210> 4872 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4872 accagattat tagacca 17 <210> 4873 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4873 acatactctg tgctgac 17 <210> 4874 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4874 ttacacatac tctgtgctga 20 <210> 4875 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4875 cttagtagtc atctcca 17 <210> 4876 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4876 acttagtagt catctcc 17 <210> 4877 <211> 17 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4877 gacttagtag tcatctc 17 <210> 4878 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4878 agcttcttgt ccagctttat 20 <210> 4879 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4879 agcttcttgt ccagctttat a 21 <210> 4880 <211> 14 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4880 tcagtcatga cttc 14 <210> 4881 <211> 15 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4881 tcagtcatga cttca 15 <210> 4882 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4882 gctgattaga gagaggtccc 20 <210> 4883 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4883 tcccatttca ggagacctgg 20 <210> 4884 <211> 15 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4884 atcagtcatg acttc 15 <210> 4885 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4885 cggtgcaagg cttaggaatt 20 <210> 4886 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4886 gcttcagtca tgacttcctt 20 <210> 4887 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4887 gcttcagtca tgacttcctt a 21 <210> 4888 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4888 agcttcagtc atgacttcct t 21 <210> 4889 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4889 tggtaatcca ctttcagagg 20 <210> 4890 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4890 tggtaatcca ctttcagagg a 21 <210> 4891 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4891 tgcttcagtc atgacttcct t 21 <210> 4892 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4892 cactgatttt tgcccaggat 20 <210> 4893 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4893 cactgatttt tgcccaggat a 21 <210> 4894 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4894 aagcttcttg tccagcttta t 21 <210> 4895 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4895 acccaattca gaaggaagga 20 <210> 4896 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4896 acccaattca gaaggaagga a 21 <210> 4897 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4897 aacccaattc agaaggaagg a 21 <210> 4898 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4898 atggtaatcc actttcagag g 21 <210> 4899 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4899 tcttggttac atgaaatccc 20 <210> 4900 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4900 tcttggttac atgaaatccc a 21 <210> 4901 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4901 attcactttc ataatgctgg 20 <210> 4902 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4902 attcactttc ataatgctgg a 21 <210> 4903 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4903 tgctccgttg gtgcttgttc 20 <210> 4904 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4904 atcttggtta catgaaatcc c 21 <210> 4905 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4905 atgcatggtg atgcttctga 20 <210> 4906 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4906 cagctttatt agggacagca 20 <210> 4907 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4907 cagctttatt agggacagca a 21 <210> 4908 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4908 acagctttat tagggacagc a 21 <210> 4909 <211> 16 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4909 ttcagtcatg acttcc 16 <210> 4910 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <220> <221> misc_feature <222> (1)..(5) <223> Bases at these positions are RNA <220> <221> misc_feature <222> (16)..(20) <223> Bases at these positions are RNA <400> 4910 gcuucagtca tgactuccuu 20 <210> 4911 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4911 tgctccgttg gtgcttgttc a 21 <210> 4912 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4912 ggacattgcc agtaatcgca a 21 <210> 4913 <211> 21 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4913 aggacattgc cagtaatcgc a 21 <210> 4914 <211> 20 <212> DNA <213> Artificial sequence <220> <223> Synthetic Oligonucleotide <400> 4914 gacatgttct tcacctcctc 20

Claims (188)

A compound comprising a modified oligonucleotide and a conjugate group, or a pharmaceutically acceptable salt thereof, comprising:
The nucleobase sequence of the modified oligonucleotide comprises 16 to 30 linked nucleosides having a nucleobase sequence comprising at least 16 contiguous nucleobases complementary to an equivalent length portion of nucleobases 1140 to 1159 of SEQ ID NO: 1 wherein the nucleobase sequence of the modified oligonucleotide is complementary to SEQ ID NO: 1 as measured throughout the modified oligonucleotide,
The conjugate group comprises the formula GalNAc ₃ -L, wherein GalNAc ₃ is of the structure

is a cell-targeting moiety having a;
L is the structure

is a conjugate linker having a, wherein the acyl group of L is attached to _{GalNAc 3 ;}
each n is independently selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, and 7,
A compound or a pharmaceutically acceptable salt thereof. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein the modified oligonucleotide comprises 20 linked nucleosides and has the nucleobase sequence of SEQ ID NO: 77. delete The compound according to claim 1, wherein the modified oligonucleotide is single-stranded or double-stranded, or a pharmaceutically acceptable salt thereof. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the modified oligonucleotide comprises at least one phosphorothioate internucleoside linkage. 6. The compound or pharmaceutically acceptable salt thereof according to claim 5, wherein the modified oligonucleotide comprises at least one phosphodiester internucleoside linkage. The compound or pharmaceutically acceptable salt thereof according to claim 6, wherein the modified oligonucleotide comprises at least 7 phosphodiester internucleoside linkages. The compound or pharmaceutical thereof according to claim 5, wherein each internucleoside linkage of the modified oligonucleotide is selected from the group consisting of a phosphodiester internucleoside linkage and a phosphorothioate internucleoside linkage. acceptable salts. The compound according to claim 8, or a pharmaceutically acceptable salt thereof, wherein each internucleoside linkage of the modified oligonucleotide is a phosphorothioate internucleoside linkage. 6. The compound of claim 5, wherein the modified oligonucleotide comprises at least one furanose comprising at least one non-naturally occurring substitution. The compound or a pharmaceutically acceptable salt thereof according to claim 10, wherein the furanose comprises a bridge connecting two ring carbon atoms. 11. The method of claim 10, wherein the furanose is 2'-O-methoxyethyl furanose, restricted ethyl furanose, 3'-fluoro-HNA furanose, and 4'-(CH ₂ ) _n -O-2' bridge A compound selected from the group consisting of furanose comprising, wherein n is 1 or 2, or a pharmaceutically acceptable salt thereof. The compound of claim 12 , wherein the at least one nucleoside comprises 5-methylcytosine, or a pharmaceutically acceptable salt thereof. 14. The method of claim 13, wherein the modified oligonucleotide is
a gap segment consisting of linked deoxynucleosides;
a 5' wing segment consisting of linked nucleosides; and
3' wing segment with linked nucleosides
comprising;
wherein said gap segment is disposed between said 5' wing segment and said 3' wing segment, wherein each nucleoside of each wing segment comprises a furanose comprising at least one non-naturally occurring substitution; or A pharmaceutically acceptable salt thereof. A compound comprising a modified oligonucleotide and a conjugate group, or a pharmaceutically acceptable salt thereof, comprising:
wherein the modified oligonucleotide comprises 20 linked nucleosides and has a nucleobase sequence comprising at least 16 contiguous nucleobases of SEQ ID NO: 77, wherein the modified oligonucleotide comprises:
a gap segment consisting of 10 linked deoxynucleosides;
5' wing segment consisting of 5 linked nucleosides;
3' wing segment with 5 linked nucleosides
comprising;
wherein the gap segment is disposed between the 5' wing segment and the 3' wing segment, wherein each nucleoside of each wing segment comprises 2'-O-methoxyethyl furanose, and wherein the modified oligonucleotide comprises: A compound or a pharmaceutically acceptable salt thereof comprising at least one phosphorothioate linkage, wherein each cytosine residue is 5-methylcytosine. A compound of the formula: or a pharmaceutically acceptable salt thereof.

The compound according to claim 14, wherein the pharmaceutically acceptable salt is a sodium salt or a potassium salt, or a pharmaceutically acceptable salt thereof. The compound according to claim 15 or 16, or a pharmaceutically acceptable salt thereof, wherein the pharmaceutically acceptable salt is a sodium salt or a potassium salt. The compound according to claim 17, wherein the conjugate group is attached to the modified oligonucleotide at the 5' end of the modified oligonucleotide, or a pharmaceutically acceptable salt thereof. 20. The method of any one of claims 1, 2, 4 to 14, 17 and 19, wherein L is

A phosphorus compound or a pharmaceutically acceptable salt thereof. A compound comprising a modified oligonucleotide and a conjugate group, or a pharmaceutically acceptable salt thereof, wherein the nucleobase sequence of the modified oligonucleotide is at least complementary to an equivalent length portion of nucleobases 1140 to 1159 of SEQ ID NO: 1 16 to 30 linked nucleosides having a nucleobase sequence comprising 16 contiguous nucleobases, wherein the nucleobase sequence of the modified oligonucleotide is SEQ ID NO: 1 as measured over the entirety of the modified oligonucleotide; complementary, wherein said conjugate group comprises the formula GalNAc ₃ -L, wherein GalNAc ₃ is of the structure

is a cell-targeting moiety having a;
L is the structure

is a conjugate linker having a, wherein the acyl group of L is attached to _{GalNAc 3 ;}
each n is independently selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, and 7, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent. containing,
A pharmaceutical composition for treating, preventing or delaying a disease associated with elevated angiopoietin-like 3 (ANGPTL3). 22. The method of claim 21, wherein the disease is obesity, diabetes, insulin resistance, atherosclerosis, dyslipidemia, lipodystrophy, coronary heart disease, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), hyperlipidemia and A pharmaceutical composition selected from the group consisting of metabolic syndrome. 23. The pharmaceutical composition of claim 22, wherein the dyslipidemia is a combination of hypertriglyceridemia and hypercholesterolemia. 23. The pharmaceutical composition of claim 22, wherein the NAFLD is hepatic steatosis. 23. The pharmaceutical composition of claim 22, wherein the diabetes is type 2 diabetes. 25. The method of any one of claims 21 to 24, wherein L is

Phosphorus pharmaceutical composition. 27. The pharmaceutical composition of claim 26, wherein the pharmaceutically acceptable salt is a sodium salt or a potassium salt. 27. The pharmaceutical composition of claim 26, wherein the pharmaceutically acceptable carrier or diluent is selected from the group consisting of sterile saline and phosphate-buffered saline (PBS). delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete